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human g6pc1 cdna  (Transomic Technologies Inc)

 
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    Transomic Technologies Inc human g6pc1 cdna
    Comparison of human <t>G6PC1</t> and G6PC2 protein expression driven by the pcDNA3.1D and pJPA5 vectors. 832/13 cells were transiently transfected with either pcDNA3.1D or pJPA5 expression vectors encoding human G6PC1 or G6PC2 with a C terminal V5 His Tag. Following transfection, cells were incubated for 18–20 h in serum-containing media. Cells were subsequently harvested and protein expression assayed by Western blotting as described in . A, G6PC expression was assessed using an anti-V5 antibody and equal protein loading was confirmed using both Ponceaus staining and measurement of actin expression. B, the conditions used to assess G6PC2 and actin expression are semiquantitative. Representative blots are shown. G6PC, glucose-6-phosphatase catalytic subunit.
    Human G6pc1 Cdna, supplied by Transomic Technologies Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+g6pc1+cdna/human+g6pc1+cdna/pmc08800118-236-1-15
    Average 90 stars, based on 1 article reviews
    human g6pc1 cdna - by Bioz Stars, 2026-10
    90/100 stars

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    1) Product Images from "Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose"

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose

    Journal: The Journal of Biological Chemistry

    doi: 10.1016/j.jbc.2021.101534

    Comparison of human G6PC1 and G6PC2 protein expression driven by the pcDNA3.1D and pJPA5 vectors. 832/13 cells were transiently transfected with either pcDNA3.1D or pJPA5 expression vectors encoding human G6PC1 or G6PC2 with a C terminal V5 His Tag. Following transfection, cells were incubated for 18–20 h in serum-containing media. Cells were subsequently harvested and protein expression assayed by Western blotting as described in . A, G6PC expression was assessed using an anti-V5 antibody and equal protein loading was confirmed using both Ponceaus staining and measurement of actin expression. B, the conditions used to assess G6PC2 and actin expression are semiquantitative. Representative blots are shown. G6PC, glucose-6-phosphatase catalytic subunit.
    Figure Legend Snippet: Comparison of human G6PC1 and G6PC2 protein expression driven by the pcDNA3.1D and pJPA5 vectors. 832/13 cells were transiently transfected with either pcDNA3.1D or pJPA5 expression vectors encoding human G6PC1 or G6PC2 with a C terminal V5 His Tag. Following transfection, cells were incubated for 18–20 h in serum-containing media. Cells were subsequently harvested and protein expression assayed by Western blotting as described in . A, G6PC expression was assessed using an anti-V5 antibody and equal protein loading was confirmed using both Ponceaus staining and measurement of actin expression. B, the conditions used to assess G6PC2 and actin expression are semiquantitative. Representative blots are shown. G6PC, glucose-6-phosphatase catalytic subunit.

    Techniques Used: Comparison, Expressing, Transfection, Incubation, Western Blot, Staining

    Conservation of amino acids between human G6PC2, mouse G6PC2, human G6PC1, and mouse G6PC1. Sequence alignment showing the conservation of AAs between human (h) and mouse (m) G6PC1 and G6PC2. Residues highlighted in yellow represent conserved AAs that are changed by a human G6PC2 SNP and where mutation in G6PC1 can cause GSD type 1a ( <xref ref-type=Table 1 ). Residues highlighted in pink represent conserved AAs that are changed by human G6PC2 SNPs but where mutation in G6PC1 has not been associated with GSD type 1a. Residues highlighted in green represent nonconserved AAs that are changed by human G6PC2 SNPs. Identities are indicated by filled circles and similarities by vertical bars. Gray boxes represent putative transmembrane domains determined using the TMHMM algorithm ( https://services.healthtech.dtu.dk/ ) . The predictions made by this program differ from earlier models ( , ). The figure shows the predicted domains for human G6PC1. G6PC1 and G6PC2 have an extended sequence motif (KXXXXXXRP-(X12-54)-PSGH-(X31-54)-SRXXXXXHXXXD) ( boxed ), similar to that found in bacterial vanadate-sensitive haloperoxidases and mammalian phosphatidic acid phosphatases, which constitutes the active site of these enzymes . G6PC1 and G6PC2 also share a C terminal ER retention sequence (KK; boxed ) . Adapted from Ref. . G6PC, glucose-6-phosphatase catalytic subunit; GSD1a, glycogen storage disease type 1a; SNP, single-nucleotide polymorphism. " title="... amino acids between human G6PC2, mouse G6PC2, human G6PC1, and mouse G6PC1. Sequence alignment showing the conservation ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Conservation of amino acids between human G6PC2, mouse G6PC2, human G6PC1, and mouse G6PC1. Sequence alignment showing the conservation of AAs between human (h) and mouse (m) G6PC1 and G6PC2. Residues highlighted in yellow represent conserved AAs that are changed by a human G6PC2 SNP and where mutation in G6PC1 can cause GSD type 1a ( Table 1 ). Residues highlighted in pink represent conserved AAs that are changed by human G6PC2 SNPs but where mutation in G6PC1 has not been associated with GSD type 1a. Residues highlighted in green represent nonconserved AAs that are changed by human G6PC2 SNPs. Identities are indicated by filled circles and similarities by vertical bars. Gray boxes represent putative transmembrane domains determined using the TMHMM algorithm ( https://services.healthtech.dtu.dk/ ) . The predictions made by this program differ from earlier models ( , ). The figure shows the predicted domains for human G6PC1. G6PC1 and G6PC2 have an extended sequence motif (KXXXXXXRP-(X12-54)-PSGH-(X31-54)-SRXXXXXHXXXD) ( boxed ), similar to that found in bacterial vanadate-sensitive haloperoxidases and mammalian phosphatidic acid phosphatases, which constitutes the active site of these enzymes . G6PC1 and G6PC2 also share a C terminal ER retention sequence (KK; boxed ) . Adapted from Ref. . G6PC, glucose-6-phosphatase catalytic subunit; GSD1a, glycogen storage disease type 1a; SNP, single-nucleotide polymorphism.

    Techniques Used: Sequencing, Mutagenesis

    Analysis of the effect of human G6PC2 SNPs that affect conserved amino acids associated with GSD1a in the context of human G6PC1
    Figure Legend Snippet: Analysis of the effect of human G6PC2 SNPs that affect conserved amino acids associated with GSD1a in the context of human G6PC1

    Techniques Used: Mutagenesis, Expressing

    Analysis of the effect of human G6PC2 SNPs that affect conserved amino acids associated with GSD1a in the context of human G6PC1, on G6PC2 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2 where a mutation is known to cause GSD1a in the context of human G6PC1 ( <xref ref-type=Table 1 ). The effect of these G6PC2 SNPs on G6PC2 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4–10. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism. " title="... associated with GSD1a in the context of human G6PC1, on G6PC2 protein expression. Human G6PC2 SNPs were ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Analysis of the effect of human G6PC2 SNPs that affect conserved amino acids associated with GSD1a in the context of human G6PC1, on G6PC2 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2 where a mutation is known to cause GSD1a in the context of human G6PC1 ( Table 1 ). The effect of these G6PC2 SNPs on G6PC2 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4–10. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism.

    Techniques Used: Expressing, Mutagenesis

    Analysis of the effect of human G6PC2 SNPs that affect amino acids that are not associated with GSD1a in the context of human G6PC1, on G6PC2 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2, which are not associated with GSD1a in the context of human G6PC1, as well as amino acids that are not conserved in G6PC1 ( <xref ref-type=Table 2 ). The effect of these G6PC2 SNPs on G6PC2 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4 to 10. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism. " title="... associated with GSD1a in the context of human G6PC1, on G6PC2 protein expression. Human G6PC2 SNPs were ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Analysis of the effect of human G6PC2 SNPs that affect amino acids that are not associated with GSD1a in the context of human G6PC1, on G6PC2 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2, which are not associated with GSD1a in the context of human G6PC1, as well as amino acids that are not conserved in G6PC1 ( Table 2 ). The effect of these G6PC2 SNPs on G6PC2 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4 to 10. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism.

    Techniques Used: Expressing

    Analysis of the effect of human G6PC2 SNPs that affect amino acids that are not associated with GSD1a in the context of human G6PC1
    Figure Legend Snippet: Analysis of the effect of human G6PC2 SNPs that affect amino acids that are not associated with GSD1a in the context of human G6PC1

    Techniques Used: Expressing

    Analysis of the suppression of glucose-stimulated fusion gene expression by G6PC2 in 832/13 cells. 832/13 cells were transiently cotransfected with the −7248/+62 G6pc1 - luciferase fusion gene (2 μg), an expression vector encoding Renilla luciferase (0.5 μg) and expression vectors (1 μg) encoding either wild type (WT), catalytically dead (D; AA 174 Ala), or variant (AA8, AA114) G6PC2. Following transfection, cells were incubated for 18 to 20 h in serum-free medium in the presence of 2 or 11 mM glucose. Cells were then harvested and luciferase activity assayed as described in . Results show mean data −/+ SD (n = 3–6) and were calculated as the ratio of firefly: Renilla luciferase activity. Statistical comparisons to 11 mM WT were made using ANOVA with a Dunnett's Multiple Comparisons post-hoc test; ∗ p < 0.05. G6PC, glucose-6-phosphatase catalytic subunit.
    Figure Legend Snippet: Analysis of the suppression of glucose-stimulated fusion gene expression by G6PC2 in 832/13 cells. 832/13 cells were transiently cotransfected with the −7248/+62 G6pc1 - luciferase fusion gene (2 μg), an expression vector encoding Renilla luciferase (0.5 μg) and expression vectors (1 μg) encoding either wild type (WT), catalytically dead (D; AA 174 Ala), or variant (AA8, AA114) G6PC2. Following transfection, cells were incubated for 18 to 20 h in serum-free medium in the presence of 2 or 11 mM glucose. Cells were then harvested and luciferase activity assayed as described in . Results show mean data −/+ SD (n = 3–6) and were calculated as the ratio of firefly: Renilla luciferase activity. Statistical comparisons to 11 mM WT were made using ANOVA with a Dunnett's Multiple Comparisons post-hoc test; ∗ p < 0.05. G6PC, glucose-6-phosphatase catalytic subunit.

    Techniques Used: Gene Expression, Luciferase, Expressing, Plasmid Preparation, Variant Assay, Transfection, Incubation, Activity Assay

    Analysis of the effect of human G6PC2 SNPs on G6PC1 protein expression. 832/13 cells were transiently transfected with pcDNA3.1D expression vectors encoding human G6PC1 with a C terminal V5 His Tag. Following transfection, cells were incubated for 18–20 h in serum-containing media. Cells were subsequently harvested and protein expression assayed by Western blotting as described in . G6PC1 expression was assessed using an anti-V5 antibody and equal protein loading was confirmed by measurement of actin expression. Representative blots are shown. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism.
    Figure Legend Snippet: Analysis of the effect of human G6PC2 SNPs on G6PC1 protein expression. 832/13 cells were transiently transfected with pcDNA3.1D expression vectors encoding human G6PC1 with a C terminal V5 His Tag. Following transfection, cells were incubated for 18–20 h in serum-containing media. Cells were subsequently harvested and protein expression assayed by Western blotting as described in . G6PC1 expression was assessed using an anti-V5 antibody and equal protein loading was confirmed by measurement of actin expression. Representative blots are shown. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism.

    Techniques Used: Expressing, Transfection, Incubation, Western Blot

    Analysis of the effect of human G6PC2 SNPs that affect conserved amino acids associated with GSD1a in the context of human G6PC1, on G6PC1 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2 where a mutation is known to cause GSD1a in the context of human G6PC1 ( <xref ref-type=Table 1 ). The effect of these G6PC2 SNPs on G6PC1 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4–8. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism. " title="... associated with GSD1a in the context of human G6PC1, on G6PC1 protein expression. Human G6PC2 SNPs were ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Analysis of the effect of human G6PC2 SNPs that affect conserved amino acids associated with GSD1a in the context of human G6PC1, on G6PC1 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2 where a mutation is known to cause GSD1a in the context of human G6PC1 ( Table 1 ). The effect of these G6PC2 SNPs on G6PC1 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4–8. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism.

    Techniques Used: Expressing, Mutagenesis

    Analysis of the effect of human G6PC2 SNPs that affect amino acids that are not associated with GSD1a in the context of human G6PC1, on G6PC1 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2, that are not associated with GSD1a in the context of human G6PC1 ( <xref ref-type=Table 2 ). The effect of these G6PC2 SNPs on G6PC1 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4–10. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism. " title="... associated with GSD1a in the context of human G6PC1, on G6PC1 protein expression. Human G6PC2 SNPs were ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Analysis of the effect of human G6PC2 SNPs that affect amino acids that are not associated with GSD1a in the context of human G6PC1, on G6PC1 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2, that are not associated with GSD1a in the context of human G6PC1 ( Table 2 ). The effect of these G6PC2 SNPs on G6PC1 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4–10. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism.

    Techniques Used: Expressing

    Comparison of the effect of specific amino acid changes on the expression of mouse and human G6PC1. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2 ( <xref ref-type=Tables 1 and ). The effect of these G6PC2 SNPs on mouse and human G6PC1 protein expression was determined as described in Fig. 3 . A representative blot is shown. G6PC, glucose-6-phosphatase catalytic subunit. " title="... changes on the expression of mouse and human G6PC1. Human G6PC2 SNPs were identified that affect AAs ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Comparison of the effect of specific amino acid changes on the expression of mouse and human G6PC1. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2 ( Tables 1 and ). The effect of these G6PC2 SNPs on mouse and human G6PC1 protein expression was determined as described in Fig. 3 . A representative blot is shown. G6PC, glucose-6-phosphatase catalytic subunit.

    Techniques Used: Comparison, Expressing

    Related Articles

    Comparison:

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose
    Article Snippet: this study, the human G6PC2 sequence designated as WT contained a leucine at AA219. .. A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies. .. This plasmid was used as the template in a PCR reaction with the following primers to

    Expressing:

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose
    Article Snippet: this study, the human G6PC2 sequence designated as WT contained a leucine at AA219. .. A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies. .. This plasmid was used as the template in a PCR reaction with the following primers to

    Transfection:

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose
    Article Snippet: this study, the human G6PC2 sequence designated as WT contained a leucine at AA219. .. A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies. .. This plasmid was used as the template in a PCR reaction with the following primers to

    Incubation:

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose
    Article Snippet: this study, the human G6PC2 sequence designated as WT contained a leucine at AA219. .. A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies. .. This plasmid was used as the template in a PCR reaction with the following primers to

    Western Blot:

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose
    Article Snippet: this study, the human G6PC2 sequence designated as WT contained a leucine at AA219. .. A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies. .. This plasmid was used as the template in a PCR reaction with the following primers to

    Staining:

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose
    Article Snippet: this study, the human G6PC2 sequence designated as WT contained a leucine at AA219. .. A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies. .. This plasmid was used as the template in a PCR reaction with the following primers to

    Sequencing:

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose
    Article Snippet: this study, the human G6PC2 sequence designated as WT contained a leucine at AA219. .. A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies. .. This plasmid was used as the template in a PCR reaction with the following primers to

    Mutagenesis:

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose
    Article Snippet: this study, the human G6PC2 sequence designated as WT contained a leucine at AA219. .. A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies. .. This plasmid was used as the template in a PCR reaction with the following primers to

    Gene Expression:

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose
    Article Snippet: this study, the human G6PC2 sequence designated as WT contained a leucine at AA219. .. A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies. .. This plasmid was used as the template in a PCR reaction with the following primers to

    Luciferase:

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose
    Article Snippet: this study, the human G6PC2 sequence designated as WT contained a leucine at AA219. .. A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies. .. This plasmid was used as the template in a PCR reaction with the following primers to

    Plasmid Preparation:

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose
    Article Snippet: this study, the human G6PC2 sequence designated as WT contained a leucine at AA219. .. A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies. .. This plasmid was used as the template in a PCR reaction with the following primers to

    Variant Assay:

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose
    Article Snippet: this study, the human G6PC2 sequence designated as WT contained a leucine at AA219. .. A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies. .. This plasmid was used as the template in a PCR reaction with the following primers to

    Activity Assay:

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose
    Article Snippet: this study, the human G6PC2 sequence designated as WT contained a leucine at AA219. .. A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies. .. This plasmid was used as the template in a PCR reaction with the following primers to



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    The 10–12-week-old heterozygous (mR83/huR83C) mice, not fasted, were treated by systemic administration of BEAM-301 and the target site sequence was analyzed one week later via next-generation sequencing (NGS). a Target site sequence for p.R83C to p.R83 correction, the p.R83 synonymous variant and the pathogenic p.Y85H bystander variant. Nucleotides on the complementary strand edited by BEAM-301 are bracketed. b Correlation of dose levels of BEAM-301 to the editing efficiency for the conversion of the p.R83C ( G6PC1 - c.247C > T ) variant to wild-type p.R83 ( G6PC1 - c.247 C ) along with the p.Y85H mutation. The n numbers for each dosage of BEAM-301 in mg/kg are listed. c Allele frequencies resulting from dosing heterozygous mice at 1.5 mg/kg (301H): p.R83C to p.R83 (correction); p.Y85H bystander mutation; Uncorrected (synonymous) editing (R83C); and Indels ( n = 5). Statistics were performed using a two-tailed unpaired T test. Data are presented as Mean values ± SEM, and individual data points for each animal are displayed. * denotes p < 0.05, ** denotes p value < 0.005.

    Journal: Nature Communications

    Article Title: Base-editing corrects metabolic abnormalities in a humanized mouse model for glycogen storage disease type-Ia

    doi: 10.1038/s41467-024-54108-1

    Figure Lengend Snippet: The 10–12-week-old heterozygous (mR83/huR83C) mice, not fasted, were treated by systemic administration of BEAM-301 and the target site sequence was analyzed one week later via next-generation sequencing (NGS). a Target site sequence for p.R83C to p.R83 correction, the p.R83 synonymous variant and the pathogenic p.Y85H bystander variant. Nucleotides on the complementary strand edited by BEAM-301 are bracketed. b Correlation of dose levels of BEAM-301 to the editing efficiency for the conversion of the p.R83C ( G6PC1 - c.247C > T ) variant to wild-type p.R83 ( G6PC1 - c.247 C ) along with the p.Y85H mutation. The n numbers for each dosage of BEAM-301 in mg/kg are listed. c Allele frequencies resulting from dosing heterozygous mice at 1.5 mg/kg (301H): p.R83C to p.R83 (correction); p.Y85H bystander mutation; Uncorrected (synonymous) editing (R83C); and Indels ( n = 5). Statistics were performed using a two-tailed unpaired T test. Data are presented as Mean values ± SEM, and individual data points for each animal are displayed. * denotes p < 0.05, ** denotes p value < 0.005.

    Article Snippet: A human cDNA encoding the open reading frame for G6PC1 - c.247C > T ( G6PC1 -R83C) were inserted into exon 1 of the mouse G6pc gene at the ATG start codon (Taconic Biosciences) in a way that created a premature STOP codon in the coding sequence of the mouse G6pc exon 1.

    Techniques: Sequencing, Next-Generation Sequencing, Variant Assay, Mutagenesis, Two Tailed Test

    Comparison of human G6PC1 and G6PC2 protein expression driven by the pcDNA3.1D and pJPA5 vectors. 832/13 cells were transiently transfected with either pcDNA3.1D or pJPA5 expression vectors encoding human G6PC1 or G6PC2 with a C terminal V5 His Tag. Following transfection, cells were incubated for 18–20 h in serum-containing media. Cells were subsequently harvested and protein expression assayed by Western blotting as described in . A, G6PC expression was assessed using an anti-V5 antibody and equal protein loading was confirmed using both Ponceaus staining and measurement of actin expression. B, the conditions used to assess G6PC2 and actin expression are semiquantitative. Representative blots are shown. G6PC, glucose-6-phosphatase catalytic subunit.

    Journal: The Journal of Biological Chemistry

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose

    doi: 10.1016/j.jbc.2021.101534

    Figure Lengend Snippet: Comparison of human G6PC1 and G6PC2 protein expression driven by the pcDNA3.1D and pJPA5 vectors. 832/13 cells were transiently transfected with either pcDNA3.1D or pJPA5 expression vectors encoding human G6PC1 or G6PC2 with a C terminal V5 His Tag. Following transfection, cells were incubated for 18–20 h in serum-containing media. Cells were subsequently harvested and protein expression assayed by Western blotting as described in . A, G6PC expression was assessed using an anti-V5 antibody and equal protein loading was confirmed using both Ponceaus staining and measurement of actin expression. B, the conditions used to assess G6PC2 and actin expression are semiquantitative. Representative blots are shown. G6PC, glucose-6-phosphatase catalytic subunit.

    Article Snippet: A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies.

    Techniques: Comparison, Expressing, Transfection, Incubation, Western Blot, Staining

    Conservation of amino acids between human G6PC2, mouse G6PC2, human G6PC1, and mouse G6PC1. Sequence alignment showing the conservation of AAs between human (h) and mouse (m) G6PC1 and G6PC2. Residues highlighted in yellow represent conserved AAs that are changed by a human G6PC2 SNP and where mutation in G6PC1 can cause GSD type 1a ( <xref ref-type=Table 1 ). Residues highlighted in pink represent conserved AAs that are changed by human G6PC2 SNPs but where mutation in G6PC1 has not been associated with GSD type 1a. Residues highlighted in green represent nonconserved AAs that are changed by human G6PC2 SNPs. Identities are indicated by filled circles and similarities by vertical bars. Gray boxes represent putative transmembrane domains determined using the TMHMM algorithm ( https://services.healthtech.dtu.dk/ ) . The predictions made by this program differ from earlier models ( , ). The figure shows the predicted domains for human G6PC1. G6PC1 and G6PC2 have an extended sequence motif (KXXXXXXRP-(X12-54)-PSGH-(X31-54)-SRXXXXXHXXXD) ( boxed ), similar to that found in bacterial vanadate-sensitive haloperoxidases and mammalian phosphatidic acid phosphatases, which constitutes the active site of these enzymes . G6PC1 and G6PC2 also share a C terminal ER retention sequence (KK; boxed ) . Adapted from Ref. . G6PC, glucose-6-phosphatase catalytic subunit; GSD1a, glycogen storage disease type 1a; SNP, single-nucleotide polymorphism. " width="100%" height="100%">

    Journal: The Journal of Biological Chemistry

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose

    doi: 10.1016/j.jbc.2021.101534

    Figure Lengend Snippet: Conservation of amino acids between human G6PC2, mouse G6PC2, human G6PC1, and mouse G6PC1. Sequence alignment showing the conservation of AAs between human (h) and mouse (m) G6PC1 and G6PC2. Residues highlighted in yellow represent conserved AAs that are changed by a human G6PC2 SNP and where mutation in G6PC1 can cause GSD type 1a ( Table 1 ). Residues highlighted in pink represent conserved AAs that are changed by human G6PC2 SNPs but where mutation in G6PC1 has not been associated with GSD type 1a. Residues highlighted in green represent nonconserved AAs that are changed by human G6PC2 SNPs. Identities are indicated by filled circles and similarities by vertical bars. Gray boxes represent putative transmembrane domains determined using the TMHMM algorithm ( https://services.healthtech.dtu.dk/ ) . The predictions made by this program differ from earlier models ( , ). The figure shows the predicted domains for human G6PC1. G6PC1 and G6PC2 have an extended sequence motif (KXXXXXXRP-(X12-54)-PSGH-(X31-54)-SRXXXXXHXXXD) ( boxed ), similar to that found in bacterial vanadate-sensitive haloperoxidases and mammalian phosphatidic acid phosphatases, which constitutes the active site of these enzymes . G6PC1 and G6PC2 also share a C terminal ER retention sequence (KK; boxed ) . Adapted from Ref. . G6PC, glucose-6-phosphatase catalytic subunit; GSD1a, glycogen storage disease type 1a; SNP, single-nucleotide polymorphism.

    Article Snippet: A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies.

    Techniques: Sequencing, Mutagenesis

    Analysis of the effect of human G6PC2 SNPs that affect conserved amino acids associated with GSD1a in the context of human G6PC1

    Journal: The Journal of Biological Chemistry

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose

    doi: 10.1016/j.jbc.2021.101534

    Figure Lengend Snippet: Analysis of the effect of human G6PC2 SNPs that affect conserved amino acids associated with GSD1a in the context of human G6PC1

    Article Snippet: A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies.

    Techniques: Mutagenesis, Expressing

    Analysis of the effect of human G6PC2 SNPs that affect conserved amino acids associated with GSD1a in the context of human G6PC1, on G6PC2 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2 where a mutation is known to cause GSD1a in the context of human G6PC1 ( <xref ref-type=Table 1 ). The effect of these G6PC2 SNPs on G6PC2 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4–10. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism. " width="100%" height="100%">

    Journal: The Journal of Biological Chemistry

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose

    doi: 10.1016/j.jbc.2021.101534

    Figure Lengend Snippet: Analysis of the effect of human G6PC2 SNPs that affect conserved amino acids associated with GSD1a in the context of human G6PC1, on G6PC2 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2 where a mutation is known to cause GSD1a in the context of human G6PC1 ( Table 1 ). The effect of these G6PC2 SNPs on G6PC2 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4–10. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism.

    Article Snippet: A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies.

    Techniques: Expressing, Mutagenesis

    Analysis of the effect of human G6PC2 SNPs that affect amino acids that are not associated with GSD1a in the context of human G6PC1, on G6PC2 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2, which are not associated with GSD1a in the context of human G6PC1, as well as amino acids that are not conserved in G6PC1 ( <xref ref-type=Table 2 ). The effect of these G6PC2 SNPs on G6PC2 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4 to 10. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism. " width="100%" height="100%">

    Journal: The Journal of Biological Chemistry

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose

    doi: 10.1016/j.jbc.2021.101534

    Figure Lengend Snippet: Analysis of the effect of human G6PC2 SNPs that affect amino acids that are not associated with GSD1a in the context of human G6PC1, on G6PC2 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2, which are not associated with GSD1a in the context of human G6PC1, as well as amino acids that are not conserved in G6PC1 ( Table 2 ). The effect of these G6PC2 SNPs on G6PC2 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4 to 10. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism.

    Article Snippet: A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies.

    Techniques: Expressing

    Analysis of the effect of human G6PC2 SNPs that affect amino acids that are not associated with GSD1a in the context of human G6PC1

    Journal: The Journal of Biological Chemistry

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose

    doi: 10.1016/j.jbc.2021.101534

    Figure Lengend Snippet: Analysis of the effect of human G6PC2 SNPs that affect amino acids that are not associated with GSD1a in the context of human G6PC1

    Article Snippet: A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies.

    Techniques: Expressing

    Analysis of the suppression of glucose-stimulated fusion gene expression by G6PC2 in 832/13 cells. 832/13 cells were transiently cotransfected with the −7248/+62 G6pc1 - luciferase fusion gene (2 μg), an expression vector encoding Renilla luciferase (0.5 μg) and expression vectors (1 μg) encoding either wild type (WT), catalytically dead (D; AA 174 Ala), or variant (AA8, AA114) G6PC2. Following transfection, cells were incubated for 18 to 20 h in serum-free medium in the presence of 2 or 11 mM glucose. Cells were then harvested and luciferase activity assayed as described in . Results show mean data −/+ SD (n = 3–6) and were calculated as the ratio of firefly: Renilla luciferase activity. Statistical comparisons to 11 mM WT were made using ANOVA with a Dunnett's Multiple Comparisons post-hoc test; ∗ p < 0.05. G6PC, glucose-6-phosphatase catalytic subunit.

    Journal: The Journal of Biological Chemistry

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose

    doi: 10.1016/j.jbc.2021.101534

    Figure Lengend Snippet: Analysis of the suppression of glucose-stimulated fusion gene expression by G6PC2 in 832/13 cells. 832/13 cells were transiently cotransfected with the −7248/+62 G6pc1 - luciferase fusion gene (2 μg), an expression vector encoding Renilla luciferase (0.5 μg) and expression vectors (1 μg) encoding either wild type (WT), catalytically dead (D; AA 174 Ala), or variant (AA8, AA114) G6PC2. Following transfection, cells were incubated for 18 to 20 h in serum-free medium in the presence of 2 or 11 mM glucose. Cells were then harvested and luciferase activity assayed as described in . Results show mean data −/+ SD (n = 3–6) and were calculated as the ratio of firefly: Renilla luciferase activity. Statistical comparisons to 11 mM WT were made using ANOVA with a Dunnett's Multiple Comparisons post-hoc test; ∗ p < 0.05. G6PC, glucose-6-phosphatase catalytic subunit.

    Article Snippet: A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies.

    Techniques: Gene Expression, Luciferase, Expressing, Plasmid Preparation, Variant Assay, Transfection, Incubation, Activity Assay

    Analysis of the effect of human G6PC2 SNPs on G6PC1 protein expression. 832/13 cells were transiently transfected with pcDNA3.1D expression vectors encoding human G6PC1 with a C terminal V5 His Tag. Following transfection, cells were incubated for 18–20 h in serum-containing media. Cells were subsequently harvested and protein expression assayed by Western blotting as described in . G6PC1 expression was assessed using an anti-V5 antibody and equal protein loading was confirmed by measurement of actin expression. Representative blots are shown. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism.

    Journal: The Journal of Biological Chemistry

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose

    doi: 10.1016/j.jbc.2021.101534

    Figure Lengend Snippet: Analysis of the effect of human G6PC2 SNPs on G6PC1 protein expression. 832/13 cells were transiently transfected with pcDNA3.1D expression vectors encoding human G6PC1 with a C terminal V5 His Tag. Following transfection, cells were incubated for 18–20 h in serum-containing media. Cells were subsequently harvested and protein expression assayed by Western blotting as described in . G6PC1 expression was assessed using an anti-V5 antibody and equal protein loading was confirmed by measurement of actin expression. Representative blots are shown. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism.

    Article Snippet: A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies.

    Techniques: Expressing, Transfection, Incubation, Western Blot

    Analysis of the effect of human G6PC2 SNPs that affect conserved amino acids associated with GSD1a in the context of human G6PC1, on G6PC1 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2 where a mutation is known to cause GSD1a in the context of human G6PC1 ( <xref ref-type=Table 1 ). The effect of these G6PC2 SNPs on G6PC1 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4–8. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism. " width="100%" height="100%">

    Journal: The Journal of Biological Chemistry

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose

    doi: 10.1016/j.jbc.2021.101534

    Figure Lengend Snippet: Analysis of the effect of human G6PC2 SNPs that affect conserved amino acids associated with GSD1a in the context of human G6PC1, on G6PC1 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2 where a mutation is known to cause GSD1a in the context of human G6PC1 ( Table 1 ). The effect of these G6PC2 SNPs on G6PC1 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4–8. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism.

    Article Snippet: A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies.

    Techniques: Expressing, Mutagenesis

    Analysis of the effect of human G6PC2 SNPs that affect amino acids that are not associated with GSD1a in the context of human G6PC1, on G6PC1 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2, that are not associated with GSD1a in the context of human G6PC1 ( <xref ref-type=Table 2 ). The effect of these G6PC2 SNPs on G6PC1 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4–10. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism. " width="100%" height="100%">

    Journal: The Journal of Biological Chemistry

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose

    doi: 10.1016/j.jbc.2021.101534

    Figure Lengend Snippet: Analysis of the effect of human G6PC2 SNPs that affect amino acids that are not associated with GSD1a in the context of human G6PC1, on G6PC1 protein expression. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2, that are not associated with GSD1a in the context of human G6PC1 ( Table 2 ). The effect of these G6PC2 SNPs on G6PC1 protein expression was determined as described in Fig. 3 . Protein expression was quantified as described in . Results show mean data −/+ SD, n = 4–10. ∗ p < 0.05 vs. WT. G6PC, glucose-6-phosphatase catalytic subunit; SNP, single-nucleotide polymorphism.

    Article Snippet: A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies.

    Techniques: Expressing

    Comparison of the effect of specific amino acid changes on the expression of mouse and human G6PC1. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2 ( <xref ref-type=Tables 1 and ). The effect of these G6PC2 SNPs on mouse and human G6PC1 protein expression was determined as described in Fig. 3 . A representative blot is shown. G6PC, glucose-6-phosphatase catalytic subunit. " width="100%" height="100%">

    Journal: The Journal of Biological Chemistry

    Article Title: Nonsynonymous single-nucleotide polymorphisms in the G6PC2 gene affect protein expression, enzyme activity, and fasting blood glucose

    doi: 10.1016/j.jbc.2021.101534

    Figure Lengend Snippet: Comparison of the effect of specific amino acid changes on the expression of mouse and human G6PC1. Human G6PC2 SNPs were identified that affect AAs conserved between mouse and human G6PC1 and mouse and human G6PC2 ( Tables 1 and ). The effect of these G6PC2 SNPs on mouse and human G6PC1 protein expression was determined as described in Fig. 3 . A representative blot is shown. G6PC, glucose-6-phosphatase catalytic subunit.

    Article Snippet: A human G6PC1 cDNA (Accession number BC130478 ; IMAGE clone number 40146509) was purchased from Transomic technologies.

    Techniques: Comparison, Expressing