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Synthego Inc crispr cas9 guide sequence
Crispr Cas9 Guide Sequence, supplied by Synthego Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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crispr cas9 guide sequence - by Bioz Stars, 2026-11
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CRISPR:

Article Title: Suidae iPSC-derived macrophages as models for investigating susceptibility and resilience to African swine fever virus
Article Snippet: .. The CRISPR/Cas9 guide sequence (Rex1_363 CTTCTTTCACTGATTTGTAT (Synthego) is conserved between domestic pigs and red river hog and directs cleavage 8 bp upstream of the REX1 stop codon. .. Ribonucleoprotein complexes (RNPs) were formed by combining 7.5 μl sgRNA (100 μM) and 1.6 μl Cas9 protein (62 μM, IDT, #1081059) for 10 min at RT.

Article Title: Fructose induces inflammatory activation in macrophages and microglia through the nutrient‐sensing ghrelin receptor
Article Snippet: 2.3 | Fructose exposure of RAW 264.7 macrophages and IMG microglia cell lines in vitro At the beginning of the treatment, the complete medium of wild type (WT) and Ghsrmutant RAW 264.7 or IMG cell lines was replaced with the corresponding reduced FBS medium (1% FBS, 1% P/S).37–40 Specifically, in the fructose- treated group, cells (3.0 × 105 live cells per well; 6- well plate) were cultured in medium supplemented with fructose (Cat# F0127, Sigma, St. Louis, MO, USA) equimolar to the glucose content in the original respective culture medium (11.1 mM fructose was supplemented in the RPMI 1640 medium for the RAW 264.7 cell line, and 17.5 mM fructose was supplemented in the StableCellTM DMEM medium for the IMG cell line). .. To exclude the possibility that the effects observed in the fructose- treated group were due to a doubling of monosaccharide concentration rather than the specific effects of fructose, we also had a glucose- treated group CRISPR guide name Guide sequence GHSR protein target Ghsr+27371816 GUGGAACGCGACGCCCAGCG aa192- 193 Ghsr−27372373 CGGCACUCGUUGGUGUCCCG aa7- 8 T A B L E 1 Single guide RNAs were designed using Synthego's algorithm and synthesized for CRISPR editing. ..

Article Title: Combined PD-L1/TGFβ blockade allows expansion and differentiation of stem cell-like CD8 T cells in immune excluded tumors.
Article Snippet: .. EMT6 IFNGR1 KO cells were generated at Synthego (Redwood City, CA) using CRISPR KO (Guide RNA sequence: UGGUAUUCCCAGCAUACGAC; Guide RNA cut location chr10:19,597,466; Exon targeted: 2). .. The loss of IFNγ response in the EMT6 IFNGR1 KO cells was validated in vitro by stimulating the cells with mouse recombinant IFNγ Nature Communications | (2023) 14:4703 11 (catalog number n 485-MI, R&D Systems,Minneapolis, MN) for 30min at 37 °C and by analyzing STAT1 phosphorylation by flow cytometry.

Sequencing:

Article Title: Suidae iPSC-derived macrophages as models for investigating susceptibility and resilience to African swine fever virus
Article Snippet: .. The CRISPR/Cas9 guide sequence (Rex1_363 CTTCTTTCACTGATTTGTAT (Synthego) is conserved between domestic pigs and red river hog and directs cleavage 8 bp upstream of the REX1 stop codon. .. Ribonucleoprotein complexes (RNPs) were formed by combining 7.5 μl sgRNA (100 μM) and 1.6 μl Cas9 protein (62 μM, IDT, #1081059) for 10 min at RT.

Article Title: Fructose induces inflammatory activation in macrophages and microglia through the nutrient‐sensing ghrelin receptor
Article Snippet: 2.3 | Fructose exposure of RAW 264.7 macrophages and IMG microglia cell lines in vitro At the beginning of the treatment, the complete medium of wild type (WT) and Ghsrmutant RAW 264.7 or IMG cell lines was replaced with the corresponding reduced FBS medium (1% FBS, 1% P/S).37–40 Specifically, in the fructose- treated group, cells (3.0 × 105 live cells per well; 6- well plate) were cultured in medium supplemented with fructose (Cat# F0127, Sigma, St. Louis, MO, USA) equimolar to the glucose content in the original respective culture medium (11.1 mM fructose was supplemented in the RPMI 1640 medium for the RAW 264.7 cell line, and 17.5 mM fructose was supplemented in the StableCellTM DMEM medium for the IMG cell line). .. To exclude the possibility that the effects observed in the fructose- treated group were due to a doubling of monosaccharide concentration rather than the specific effects of fructose, we also had a glucose- treated group CRISPR guide name Guide sequence GHSR protein target Ghsr+27371816 GUGGAACGCGACGCCCAGCG aa192- 193 Ghsr−27372373 CGGCACUCGUUGGUGUCCCG aa7- 8 T A B L E 1 Single guide RNAs were designed using Synthego's algorithm and synthesized for CRISPR editing. ..

Article Title: Combined PD-L1/TGFβ blockade allows expansion and differentiation of stem cell-like CD8 T cells in immune excluded tumors.
Article Snippet: .. EMT6 IFNGR1 KO cells were generated at Synthego (Redwood City, CA) using CRISPR KO (Guide RNA sequence: UGGUAUUCCCAGCAUACGAC; Guide RNA cut location chr10:19,597,466; Exon targeted: 2). .. The loss of IFNγ response in the EMT6 IFNGR1 KO cells was validated in vitro by stimulating the cells with mouse recombinant IFNγ Nature Communications | (2023) 14:4703 11 (catalog number n 485-MI, R&D Systems,Minneapolis, MN) for 30min at 37 °C and by analyzing STAT1 phosphorylation by flow cytometry.

other:

Article Title: Downregulation of the MARC1 p.A165 risk allele reduces hepatocyte lipid content by increasing beta-oxidation
Article Snippet: MARC1 knock out stable cell (HepG2) generation MARC1 knock out (KO) stable cell pool was created using commercial service (Synthego’s Engineered Immortalized Cell Pools) with high-quality chemically modified synthetic single guide RNA (sgRNA) (CRISPR/Cas9 target sequence: TCTCAGTGCAGCCTACACAA, PAM sequence: AGG).

Article Title: Downregulation of the MARC1 p.A165 risk allele reduces hepatocyte lipid content by increasing beta-oxidation.
Article Snippet: MARC1 knock out stable cell (HepG2) generation: MARC1 knock out (KO) stable cell pool was created using commercial service (Synthego’s Engineered Immortalized Cell Pools) with high-quality chemically modified synthetic single guide RNA (sgRNA) (CRISPR/Cas9 target sequence: TCTCAGTGCAGCCTACACAA, PAM sequence: AGG).

Article Title: Glycogen synthase kinase 3 inhibition controls Mycobacterium tuberculosis infection.
Article Snippet: Oligonucleotides siRNA targeting sequence: Scrambled (Negative Control DsiRNA) IDT Cat# 51-01-14-04 siRNA targeting sequence: GSK3b IDT hs.Ri.GSK3B.13.3 siRNA targeting sequence: GSK3a IDT hs.Ri.GSK3A.13.2 GSK3a CRISPR Guide Sequence: GCCUAGAGUGGCUACGACUG Synthego N/A GSK3b CRISPR Guide Sequence: CGUUAUUUCUUCUACUCCAG Synthego N/A GSK3a CRISPR Sequencing For primer: CCTTCTCTTCTGGCCCATGG Synthego N/A GSK3a CRISPR Sequencing Rev primer: CAACGAGCTTCCTGCAGAGA Synthego N/A GSK3b CRISPR Sequencing For primer: TTCAGAAGCATCTTGTTCAATATG Synthego N/A GSK3b CRISPR Sequencing Rev primer: CAAGTGTTTCGGAATTTTTCC Synthego N/A GSK3b CRISPR Sequencing Rev primer: CAAGTGTTTCGGAATTTTTCC Synthego N/A Primers for qPCR, see Table S4 This paper N/A (Continued on next page) iScience 27, 110555, August 16, 2024 15

Article Title: Glycogen synthase kinase 3 inhibition controls Mycobacterium tuberculosis infection
Article Snippet: GSK3β CRISPR Guide Sequence: CGUUAUUUCUUCUACUCCAG , Synthego , N/A.

Article Title: Glycogen synthase kinase 3 inhibition controls Mycobacterium tuberculosis infection
Article Snippet: GSK3α CRISPR Guide Sequence: GCCUAGAGUGGCUACGACUG , Synthego , N/A.

Concentration Assay:

Article Title: Fructose induces inflammatory activation in macrophages and microglia through the nutrient‐sensing ghrelin receptor
Article Snippet: 2.3 | Fructose exposure of RAW 264.7 macrophages and IMG microglia cell lines in vitro At the beginning of the treatment, the complete medium of wild type (WT) and Ghsrmutant RAW 264.7 or IMG cell lines was replaced with the corresponding reduced FBS medium (1% FBS, 1% P/S).37–40 Specifically, in the fructose- treated group, cells (3.0 × 105 live cells per well; 6- well plate) were cultured in medium supplemented with fructose (Cat# F0127, Sigma, St. Louis, MO, USA) equimolar to the glucose content in the original respective culture medium (11.1 mM fructose was supplemented in the RPMI 1640 medium for the RAW 264.7 cell line, and 17.5 mM fructose was supplemented in the StableCellTM DMEM medium for the IMG cell line). .. To exclude the possibility that the effects observed in the fructose- treated group were due to a doubling of monosaccharide concentration rather than the specific effects of fructose, we also had a glucose- treated group CRISPR guide name Guide sequence GHSR protein target Ghsr+27371816 GUGGAACGCGACGCCCAGCG aa192- 193 Ghsr−27372373 CGGCACUCGUUGGUGUCCCG aa7- 8 T A B L E 1 Single guide RNAs were designed using Synthego's algorithm and synthesized for CRISPR editing. ..

Synthesized:

Article Title: Fructose induces inflammatory activation in macrophages and microglia through the nutrient‐sensing ghrelin receptor
Article Snippet: 2.3 | Fructose exposure of RAW 264.7 macrophages and IMG microglia cell lines in vitro At the beginning of the treatment, the complete medium of wild type (WT) and Ghsrmutant RAW 264.7 or IMG cell lines was replaced with the corresponding reduced FBS medium (1% FBS, 1% P/S).37–40 Specifically, in the fructose- treated group, cells (3.0 × 105 live cells per well; 6- well plate) were cultured in medium supplemented with fructose (Cat# F0127, Sigma, St. Louis, MO, USA) equimolar to the glucose content in the original respective culture medium (11.1 mM fructose was supplemented in the RPMI 1640 medium for the RAW 264.7 cell line, and 17.5 mM fructose was supplemented in the StableCellTM DMEM medium for the IMG cell line). .. To exclude the possibility that the effects observed in the fructose- treated group were due to a doubling of monosaccharide concentration rather than the specific effects of fructose, we also had a glucose- treated group CRISPR guide name Guide sequence GHSR protein target Ghsr+27371816 GUGGAACGCGACGCCCAGCG aa192- 193 Ghsr−27372373 CGGCACUCGUUGGUGUCCCG aa7- 8 T A B L E 1 Single guide RNAs were designed using Synthego's algorithm and synthesized for CRISPR editing. ..

Generated:

Article Title: Combined PD-L1/TGFβ blockade allows expansion and differentiation of stem cell-like CD8 T cells in immune excluded tumors.
Article Snippet: .. EMT6 IFNGR1 KO cells were generated at Synthego (Redwood City, CA) using CRISPR KO (Guide RNA sequence: UGGUAUUCCCAGCAUACGAC; Guide RNA cut location chr10:19,597,466; Exon targeted: 2). .. The loss of IFNγ response in the EMT6 IFNGR1 KO cells was validated in vitro by stimulating the cells with mouse recombinant IFNγ Nature Communications | (2023) 14:4703 11 (catalog number n 485-MI, R&D Systems,Minneapolis, MN) for 30min at 37 °C and by analyzing STAT1 phosphorylation by flow cytometry.



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CRISPR sgRNA/Cas9 complexes were injected into single-cell embryos that were then treated with L-thyroxine (0.6 µM) at 3 dpf and mestranol (25 µM) at 4 dpf. Statistical analyses were performed using proportional odds logistic regression (polr) models with MASS and emmeans libraries in R (v4.3.3). ****: p < 0.0001, ns: not significant. wt: wild-type (uninjected fabp-fgb-egfp control group), - thr : combined knockdown of all thyroid hormone receptor paralogs thraa , thrab , and thrb . Magenta borders indicate mestranol treated groups. All data were collected by an observer blinded to the condition. Sample sizes for each group, corresponding to the thrombosis grades, are indicated within their respective stack.

Journal: bioRxiv

Article Title: FDA-approved drug repurposing in zebrafish identifies thyroid hormone and other compounds as potential antithrombotics

doi: 10.1101/2025.09.23.677920

Figure Lengend Snippet: CRISPR sgRNA/Cas9 complexes were injected into single-cell embryos that were then treated with L-thyroxine (0.6 µM) at 3 dpf and mestranol (25 µM) at 4 dpf. Statistical analyses were performed using proportional odds logistic regression (polr) models with MASS and emmeans libraries in R (v4.3.3). ****: p < 0.0001, ns: not significant. wt: wild-type (uninjected fabp-fgb-egfp control group), - thr : combined knockdown of all thyroid hormone receptor paralogs thraa , thrab , and thrb . Magenta borders indicate mestranol treated groups. All data were collected by an observer blinded to the condition. Sample sizes for each group, corresponding to the thrombosis grades, are indicated within their respective stack.

Article Snippet: CRISPR single-guide RNA (sgRNA) sequences targeting early exons were designed with the Knockout Guide Design Tool ( synthego.com ).

Techniques: CRISPR, Injection, Control, Knockdown

CRISPR sgRNA/Cas9 complexes were injected into single-cell embryos that were then treated with tiratricol (0.1 µM) at 3 dpf and mestranol (25 µM) at 4 dpf. Statistical analyses were performed using proportional odds logistic regression (polr) models with MASS and emmeans libraries in R (v4.3.3). To prevent infinitesimal values that would be generated by the regression models, each thrombosis grade group was incremented by one count across all treatment groups. ****: p < 0.0001, ***: p < 0.001. wt: wild-type (uninjected fabp-fgb-egfp control group), - thr : combined knockdown of all thyroid hormone receptor paralogs thraa , thrab , and thrb . Magenta borders indicate mestranol treated groups. All data were collected by an observer blinded to the condition. Sample sizes for each group, corresponding to the thrombosis grades, are indicated within their respective stack.

Journal: bioRxiv

Article Title: FDA-approved drug repurposing in zebrafish identifies thyroid hormone and other compounds as potential antithrombotics

doi: 10.1101/2025.09.23.677920

Figure Lengend Snippet: CRISPR sgRNA/Cas9 complexes were injected into single-cell embryos that were then treated with tiratricol (0.1 µM) at 3 dpf and mestranol (25 µM) at 4 dpf. Statistical analyses were performed using proportional odds logistic regression (polr) models with MASS and emmeans libraries in R (v4.3.3). To prevent infinitesimal values that would be generated by the regression models, each thrombosis grade group was incremented by one count across all treatment groups. ****: p < 0.0001, ***: p < 0.001. wt: wild-type (uninjected fabp-fgb-egfp control group), - thr : combined knockdown of all thyroid hormone receptor paralogs thraa , thrab , and thrb . Magenta borders indicate mestranol treated groups. All data were collected by an observer blinded to the condition. Sample sizes for each group, corresponding to the thrombosis grades, are indicated within their respective stack.

Article Snippet: CRISPR single-guide RNA (sgRNA) sequences targeting early exons were designed with the Knockout Guide Design Tool ( synthego.com ).

Techniques: CRISPR, Injection, Generated, Control, Knockdown

CRISPR sgRNA/Cas9 complexes were injected into single-cell embryos that were then treated with L-thyroxine (0.6 µM) at 3 dpf and observed at 5 dpf. Thrombosis grades were collected simultaneously with the TTO evaluations, ensuring consistent sample use as presented in . Statistical analyses were performed using proportional odds logistic regression ( polr ) models with MASS and emmeans libraries in R (v4.3.3). ****: p < 0.0001, *: p < 0.05. - proc : genetic thrombosis line ( proc −/− ::fabp-fgb-egfp control group), - thr : combined knockdown of all thyroid hormone receptor paralogs thraa , thrab , and thrb . All data were collected by an observer blinded to the condition. Sample sizes for each group, corresponding to the thrombosis grades, are indicated within their respective stack.

Journal: bioRxiv

Article Title: FDA-approved drug repurposing in zebrafish identifies thyroid hormone and other compounds as potential antithrombotics

doi: 10.1101/2025.09.23.677920

Figure Lengend Snippet: CRISPR sgRNA/Cas9 complexes were injected into single-cell embryos that were then treated with L-thyroxine (0.6 µM) at 3 dpf and observed at 5 dpf. Thrombosis grades were collected simultaneously with the TTO evaluations, ensuring consistent sample use as presented in . Statistical analyses were performed using proportional odds logistic regression ( polr ) models with MASS and emmeans libraries in R (v4.3.3). ****: p < 0.0001, *: p < 0.05. - proc : genetic thrombosis line ( proc −/− ::fabp-fgb-egfp control group), - thr : combined knockdown of all thyroid hormone receptor paralogs thraa , thrab , and thrb . All data were collected by an observer blinded to the condition. Sample sizes for each group, corresponding to the thrombosis grades, are indicated within their respective stack.

Article Snippet: CRISPR single-guide RNA (sgRNA) sequences targeting early exons were designed with the Knockout Guide Design Tool ( synthego.com ).

Techniques: CRISPR, Injection, Control, Knockdown

CRISPR sgRNA/Cas9 complexes were injected into single-cell embryos that were then treated with L-thyroxine (0.6 µM) at 3 dpf, and mestranol (25 µM) at 4 dpf, followed by assessment of laser-mediated endothelial injury at 5 dpf. Increased TTO observed in wild-type embryos treated with thyroid hormone was diminished upon thr knockdown, indicating involvement of thyroid receptor signaling. Increased TTO levels were still present in the L-thyroxine-mestranol combination groups. ****: p<0.0001, ***: p<0.001, **: p<0.01, *: p<0.05, ns: not significant. wt: wild-type (uninjected fabp-fgb-egfp control group), - thr : combined knockdown of all thyroid hormone receptor paralogs thraa , thrab , and thrb . All data were collected by an observer blinded to the condition.

Journal: bioRxiv

Article Title: FDA-approved drug repurposing in zebrafish identifies thyroid hormone and other compounds as potential antithrombotics

doi: 10.1101/2025.09.23.677920

Figure Lengend Snippet: CRISPR sgRNA/Cas9 complexes were injected into single-cell embryos that were then treated with L-thyroxine (0.6 µM) at 3 dpf, and mestranol (25 µM) at 4 dpf, followed by assessment of laser-mediated endothelial injury at 5 dpf. Increased TTO observed in wild-type embryos treated with thyroid hormone was diminished upon thr knockdown, indicating involvement of thyroid receptor signaling. Increased TTO levels were still present in the L-thyroxine-mestranol combination groups. ****: p<0.0001, ***: p<0.001, **: p<0.01, *: p<0.05, ns: not significant. wt: wild-type (uninjected fabp-fgb-egfp control group), - thr : combined knockdown of all thyroid hormone receptor paralogs thraa , thrab , and thrb . All data were collected by an observer blinded to the condition.

Article Snippet: CRISPR single-guide RNA (sgRNA) sequences targeting early exons were designed with the Knockout Guide Design Tool ( synthego.com ).

Techniques: CRISPR, Injection, Knockdown, Control

After CRISPR-mediated individual knockdown of each canonical thyroid hormone receptor (- thraa , - thrab , and - thrb ) or combination (- thr ), embryos were co-treated with L-thyroxine (3 µM) and mestranol (25 µM) at 4 dpf and evaluated 24 hours later. Magenta borders indicate mestranol treated groups. ****: p < 0.0001, *: p < 0.05, ns: not significant. All data were collected by an observer blinded to the condition. Sample sizes for each group, corresponding to the thrombosis grades, are indicated within their respective stack.

Journal: bioRxiv

Article Title: FDA-approved drug repurposing in zebrafish identifies thyroid hormone and other compounds as potential antithrombotics

doi: 10.1101/2025.09.23.677920

Figure Lengend Snippet: After CRISPR-mediated individual knockdown of each canonical thyroid hormone receptor (- thraa , - thrab , and - thrb ) or combination (- thr ), embryos were co-treated with L-thyroxine (3 µM) and mestranol (25 µM) at 4 dpf and evaluated 24 hours later. Magenta borders indicate mestranol treated groups. ****: p < 0.0001, *: p < 0.05, ns: not significant. All data were collected by an observer blinded to the condition. Sample sizes for each group, corresponding to the thrombosis grades, are indicated within their respective stack.

Article Snippet: CRISPR single-guide RNA (sgRNA) sequences targeting early exons were designed with the Knockout Guide Design Tool ( synthego.com ).

Techniques: CRISPR, Knockdown