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cas9 coexpression vector pldcn  (Addgene inc)


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    Addgene inc cas9 coexpression vector pldcn
    Generation of LPG2 knockout using <t>CRISPR/Cas9.</t> (A) Western blot analysis of L. infantum promastigotes expressing Cas9 (gRNA440 and gRNA516). (B) Growth curves of L. infantum wild-type (WT) and L. infantum -Cas9 (gRNA440 and gRNA516) promastigotes. (C) Agglutination assay using the CA7AE monoclonal antibody and associated growth curves. (D) Agglutination assay using Ricin 120 lectin and associated growth curves, demonstrating the selection of Δ lpg2 . The arrows indicate the time points where antibody (CA7AE) or lectin (Ricin-120) were added to the cultures, exemplifying the typical agglutination results observed.
    Cas9 Coexpression Vector Pldcn, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cas9 coexpression vector pldcn/product/Addgene inc
    Average 91 stars, based on 4 article reviews
    cas9 coexpression vector pldcn - by Bioz Stars, 2026-02
    91/100 stars

    Images

    1) Product Images from "LPG2 Gene Duplication in Leishmania infantum : A Case for CRISPR-Cas9 Gene Editing"

    Article Title: LPG2 Gene Duplication in Leishmania infantum : A Case for CRISPR-Cas9 Gene Editing

    Journal: Frontiers in Cellular and Infection Microbiology

    doi: 10.3389/fcimb.2020.00408

    Generation of LPG2 knockout using CRISPR/Cas9. (A) Western blot analysis of L. infantum promastigotes expressing Cas9 (gRNA440 and gRNA516). (B) Growth curves of L. infantum wild-type (WT) and L. infantum -Cas9 (gRNA440 and gRNA516) promastigotes. (C) Agglutination assay using the CA7AE monoclonal antibody and associated growth curves. (D) Agglutination assay using Ricin 120 lectin and associated growth curves, demonstrating the selection of Δ lpg2 . The arrows indicate the time points where antibody (CA7AE) or lectin (Ricin-120) were added to the cultures, exemplifying the typical agglutination results observed.
    Figure Legend Snippet: Generation of LPG2 knockout using CRISPR/Cas9. (A) Western blot analysis of L. infantum promastigotes expressing Cas9 (gRNA440 and gRNA516). (B) Growth curves of L. infantum wild-type (WT) and L. infantum -Cas9 (gRNA440 and gRNA516) promastigotes. (C) Agglutination assay using the CA7AE monoclonal antibody and associated growth curves. (D) Agglutination assay using Ricin 120 lectin and associated growth curves, demonstrating the selection of Δ lpg2 . The arrows indicate the time points where antibody (CA7AE) or lectin (Ricin-120) were added to the cultures, exemplifying the typical agglutination results observed.

    Techniques Used: Knock-Out, CRISPR, Western Blot, Expressing, Agglutination, Selection

    Molecular characterization of Δ lpg2 and reduced virulence phenotype. (A) Chromatogram and translated sequence showing the region of the LPG2 gene in which the precise insertion of a stop codon (denoted by an *) occurred by homologous recombination at the cleavage site of the Cas9 enzyme (nucleotides in red). The oligodonor sequence is underlined and the gRNA440 sequence is highlighted in blue. (B) Western blot analysis of the expression of LPG and PPGs in L. infantum promastigotes WT and Δ lpg2 clones. (C) Confocal immunofluorescence analysis of WT and Δ lpg2 (clone G6) parasites. Late log-phase promastigotes were adhered on Poly-L-Lysine-coated glass coverslips, fixed and incubated with the CA7AE antibody to visualize LPG and other Gal(β1,4)Man(α1-PO4) repeating unit-containing PGs (green). Differential interference contrast (DIC) for Δ lpg2 is shown in the upper left panel. Scale bar: 10 μm. (D) Axenic growth curves of late log-phase promastigotes of L. infantum wild-type (WT) and clone G6 Δ lpg2 , each point represents mean and SE. Data are representative of at least three independent assays and were collected in triplicate for each experimental condition. (E) Area under the curve (AUC) analysis of growth curves presented in (D) , *** p < 0.01. (F) Reduced virulence of Δ lpg2 parasites in human neutrophils. Human neutrophils were infected with L. infantum Ba262 wild-type and Δ lpg2 promastigotes for 3 h. Numbers of viable promastigotes shown after 24 h, with each point on the graph representing the cells from a health donor. Statistical differences were evaluated using the Student t- test, **** p < 0.001.
    Figure Legend Snippet: Molecular characterization of Δ lpg2 and reduced virulence phenotype. (A) Chromatogram and translated sequence showing the region of the LPG2 gene in which the precise insertion of a stop codon (denoted by an *) occurred by homologous recombination at the cleavage site of the Cas9 enzyme (nucleotides in red). The oligodonor sequence is underlined and the gRNA440 sequence is highlighted in blue. (B) Western blot analysis of the expression of LPG and PPGs in L. infantum promastigotes WT and Δ lpg2 clones. (C) Confocal immunofluorescence analysis of WT and Δ lpg2 (clone G6) parasites. Late log-phase promastigotes were adhered on Poly-L-Lysine-coated glass coverslips, fixed and incubated with the CA7AE antibody to visualize LPG and other Gal(β1,4)Man(α1-PO4) repeating unit-containing PGs (green). Differential interference contrast (DIC) for Δ lpg2 is shown in the upper left panel. Scale bar: 10 μm. (D) Axenic growth curves of late log-phase promastigotes of L. infantum wild-type (WT) and clone G6 Δ lpg2 , each point represents mean and SE. Data are representative of at least three independent assays and were collected in triplicate for each experimental condition. (E) Area under the curve (AUC) analysis of growth curves presented in (D) , *** p < 0.01. (F) Reduced virulence of Δ lpg2 parasites in human neutrophils. Human neutrophils were infected with L. infantum Ba262 wild-type and Δ lpg2 promastigotes for 3 h. Numbers of viable promastigotes shown after 24 h, with each point on the graph representing the cells from a health donor. Statistical differences were evaluated using the Student t- test, **** p < 0.001.

    Techniques Used: Sequencing, Homologous Recombination, Western Blot, Expressing, Clone Assay, Immunofluorescence, Incubation, Infection



    Similar Products

    cas9 coexpression vector pldcn  (Addgene inc)
    91
    Addgene inc cas9 coexpression vector pldcn
    Generation of LPG2 knockout using <t>CRISPR/Cas9.</t> (A) Western blot analysis of L. infantum promastigotes expressing Cas9 (gRNA440 and gRNA516). (B) Growth curves of L. infantum wild-type (WT) and L. infantum -Cas9 (gRNA440 and gRNA516) promastigotes. (C) Agglutination assay using the CA7AE monoclonal antibody and associated growth curves. (D) Agglutination assay using Ricin 120 lectin and associated growth curves, demonstrating the selection of Δ lpg2 . The arrows indicate the time points where antibody (CA7AE) or lectin (Ricin-120) were added to the cultures, exemplifying the typical agglutination results observed.
    Cas9 Coexpression Vector Pldcn, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cas9 coexpression vector pldcn/product/Addgene inc
    Average 91 stars, based on 1 article reviews
    cas9 coexpression vector pldcn - by Bioz Stars, 2026-02
    91/100 stars
    		  Buy from Supplier

    Image Search Results


    Generation of LPG2 knockout using CRISPR/Cas9. (A) Western blot analysis of L. infantum promastigotes expressing Cas9 (gRNA440 and gRNA516). (B) Growth curves of L. infantum wild-type (WT) and L. infantum -Cas9 (gRNA440 and gRNA516) promastigotes. (C) Agglutination assay using the CA7AE monoclonal antibody and associated growth curves. (D) Agglutination assay using Ricin 120 lectin and associated growth curves, demonstrating the selection of Δ lpg2 . The arrows indicate the time points where antibody (CA7AE) or lectin (Ricin-120) were added to the cultures, exemplifying the typical agglutination results observed.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: LPG2 Gene Duplication in Leishmania infantum : A Case for CRISPR-Cas9 Gene Editing

    doi: 10.3389/fcimb.2020.00408

    Figure Lengend Snippet: Generation of LPG2 knockout using CRISPR/Cas9. (A) Western blot analysis of L. infantum promastigotes expressing Cas9 (gRNA440 and gRNA516). (B) Growth curves of L. infantum wild-type (WT) and L. infantum -Cas9 (gRNA440 and gRNA516) promastigotes. (C) Agglutination assay using the CA7AE monoclonal antibody and associated growth curves. (D) Agglutination assay using Ricin 120 lectin and associated growth curves, demonstrating the selection of Δ lpg2 . The arrows indicate the time points where antibody (CA7AE) or lectin (Ricin-120) were added to the cultures, exemplifying the typical agglutination results observed.

    Article Snippet: The annealed guide sequence was then cloned into the gRNA and Cas9 coexpression vector (pLdCN) (Addgene plasmid #84290; http://n2t.net/addgene:84290 ; RRID:Addgene_84290 ) previously digested with Bbs I.

    Techniques: Knock-Out, CRISPR, Western Blot, Expressing, Agglutination, Selection

    Molecular characterization of Δ lpg2 and reduced virulence phenotype. (A) Chromatogram and translated sequence showing the region of the LPG2 gene in which the precise insertion of a stop codon (denoted by an *) occurred by homologous recombination at the cleavage site of the Cas9 enzyme (nucleotides in red). The oligodonor sequence is underlined and the gRNA440 sequence is highlighted in blue. (B) Western blot analysis of the expression of LPG and PPGs in L. infantum promastigotes WT and Δ lpg2 clones. (C) Confocal immunofluorescence analysis of WT and Δ lpg2 (clone G6) parasites. Late log-phase promastigotes were adhered on Poly-L-Lysine-coated glass coverslips, fixed and incubated with the CA7AE antibody to visualize LPG and other Gal(β1,4)Man(α1-PO4) repeating unit-containing PGs (green). Differential interference contrast (DIC) for Δ lpg2 is shown in the upper left panel. Scale bar: 10 μm. (D) Axenic growth curves of late log-phase promastigotes of L. infantum wild-type (WT) and clone G6 Δ lpg2 , each point represents mean and SE. Data are representative of at least three independent assays and were collected in triplicate for each experimental condition. (E) Area under the curve (AUC) analysis of growth curves presented in (D) , *** p < 0.01. (F) Reduced virulence of Δ lpg2 parasites in human neutrophils. Human neutrophils were infected with L. infantum Ba262 wild-type and Δ lpg2 promastigotes for 3 h. Numbers of viable promastigotes shown after 24 h, with each point on the graph representing the cells from a health donor. Statistical differences were evaluated using the Student t- test, **** p < 0.001.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: LPG2 Gene Duplication in Leishmania infantum : A Case for CRISPR-Cas9 Gene Editing

    doi: 10.3389/fcimb.2020.00408

    Figure Lengend Snippet: Molecular characterization of Δ lpg2 and reduced virulence phenotype. (A) Chromatogram and translated sequence showing the region of the LPG2 gene in which the precise insertion of a stop codon (denoted by an *) occurred by homologous recombination at the cleavage site of the Cas9 enzyme (nucleotides in red). The oligodonor sequence is underlined and the gRNA440 sequence is highlighted in blue. (B) Western blot analysis of the expression of LPG and PPGs in L. infantum promastigotes WT and Δ lpg2 clones. (C) Confocal immunofluorescence analysis of WT and Δ lpg2 (clone G6) parasites. Late log-phase promastigotes were adhered on Poly-L-Lysine-coated glass coverslips, fixed and incubated with the CA7AE antibody to visualize LPG and other Gal(β1,4)Man(α1-PO4) repeating unit-containing PGs (green). Differential interference contrast (DIC) for Δ lpg2 is shown in the upper left panel. Scale bar: 10 μm. (D) Axenic growth curves of late log-phase promastigotes of L. infantum wild-type (WT) and clone G6 Δ lpg2 , each point represents mean and SE. Data are representative of at least three independent assays and were collected in triplicate for each experimental condition. (E) Area under the curve (AUC) analysis of growth curves presented in (D) , *** p < 0.01. (F) Reduced virulence of Δ lpg2 parasites in human neutrophils. Human neutrophils were infected with L. infantum Ba262 wild-type and Δ lpg2 promastigotes for 3 h. Numbers of viable promastigotes shown after 24 h, with each point on the graph representing the cells from a health donor. Statistical differences were evaluated using the Student t- test, **** p < 0.001.

    Article Snippet: The annealed guide sequence was then cloned into the gRNA and Cas9 coexpression vector (pLdCN) (Addgene plasmid #84290; http://n2t.net/addgene:84290 ; RRID:Addgene_84290 ) previously digested with Bbs I.

    Techniques: Sequencing, Homologous Recombination, Western Blot, Expressing, Clone Assay, Immunofluorescence, Incubation, Infection