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a) An NPC donor line heterozygous for index SNP (rs11709284) was transduced with lentivirus carrying the indicated gRNA, and dSpCas9:KRAB (CRISPRi) or <t>dSpCas9:VP64</t> <t>(CRISPRa).</t> Cells were incubated for four days, followed by an eight hour treatment with 1.5 mM LiCl. GNL3 expression was analyzed by RT-qPCR. Data normalized to neg. control gRNA (non-targeting, dashed line). n=4 qPCR replicates, significance determined by two sample t-test. * p<0.05, ** p<0.01, *** p<0.001. b) Proportion of NPCs in S-phase expressing indicated dSpCas9 and gRNAs at baseline, analyzed by high content imaging, as in . n=7 wells per condition, nine tiled images per well, significance determined by two sample t-test. c) Change in the proportion of NPCs in S-phase in response to 1.5mM LiCl, while expressing indicated dSpCas9 and gRNAs. n=7 wells per condition, significance determined by two sample t-test.
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a) An NPC donor line heterozygous for index SNP (rs11709284) was transduced with lentivirus carrying the indicated gRNA, and dSpCas9:KRAB (CRISPRi) or <t>dSpCas9:VP64</t> <t>(CRISPRa).</t> Cells were incubated for four days, followed by an eight hour treatment with 1.5 mM LiCl. GNL3 expression was analyzed by RT-qPCR. Data normalized to neg. control gRNA (non-targeting, dashed line). n=4 qPCR replicates, significance determined by two sample t-test. * p<0.05, ** p<0.01, *** p<0.001. b) Proportion of NPCs in S-phase expressing indicated dSpCas9 and gRNAs at baseline, analyzed by high content imaging, as in . n=7 wells per condition, nine tiled images per well, significance determined by two sample t-test. c) Change in the proportion of NPCs in S-phase in response to 1.5mM LiCl, while expressing indicated dSpCas9 and gRNAs. n=7 wells per condition, significance determined by two sample t-test.
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a) An NPC donor line heterozygous for index SNP (rs11709284) was transduced with lentivirus carrying the indicated gRNA, and dSpCas9:KRAB (CRISPRi) or <t>dSpCas9:VP64</t> <t>(CRISPRa).</t> Cells were incubated for four days, followed by an eight hour treatment with 1.5 mM LiCl. GNL3 expression was analyzed by RT-qPCR. Data normalized to neg. control gRNA (non-targeting, dashed line). n=4 qPCR replicates, significance determined by two sample t-test. * p<0.05, ** p<0.01, *** p<0.001. b) Proportion of NPCs in S-phase expressing indicated dSpCas9 and gRNAs at baseline, analyzed by high content imaging, as in . n=7 wells per condition, nine tiled images per well, significance determined by two sample t-test. c) Change in the proportion of NPCs in S-phase in response to 1.5mM LiCl, while expressing indicated dSpCas9 and gRNAs. n=7 wells per condition, significance determined by two sample t-test.
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Average 91 stars, based on 1 article reviews
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anti human tdg antibody - by Bioz Stars, 2025-01
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a) An NPC donor line heterozygous for index SNP (rs11709284) was transduced with lentivirus carrying the indicated gRNA, and dSpCas9:KRAB (CRISPRi) or dSpCas9:VP64 (CRISPRa). Cells were incubated for four days, followed by an eight hour treatment with 1.5 mM LiCl. GNL3 expression was analyzed by RT-qPCR. Data normalized to neg. control gRNA (non-targeting, dashed line). n=4 qPCR replicates, significance determined by two sample t-test. * p<0.05, ** p<0.01, *** p<0.001. b) Proportion of NPCs in S-phase expressing indicated dSpCas9 and gRNAs at baseline, analyzed by high content imaging, as in . n=7 wells per condition, nine tiled images per well, significance determined by two sample t-test. c) Change in the proportion of NPCs in S-phase in response to 1.5mM LiCl, while expressing indicated dSpCas9 and gRNAs. n=7 wells per condition, significance determined by two sample t-test.

Journal: bioRxiv

Article Title: Cellular genome wide association study identifies common genetic variation influencing lithium induced neural progenitor proliferation

doi: 10.1101/2022.01.31.478307

Figure Lengend Snippet: a) An NPC donor line heterozygous for index SNP (rs11709284) was transduced with lentivirus carrying the indicated gRNA, and dSpCas9:KRAB (CRISPRi) or dSpCas9:VP64 (CRISPRa). Cells were incubated for four days, followed by an eight hour treatment with 1.5 mM LiCl. GNL3 expression was analyzed by RT-qPCR. Data normalized to neg. control gRNA (non-targeting, dashed line). n=4 qPCR replicates, significance determined by two sample t-test. * p<0.05, ** p<0.01, *** p<0.001. b) Proportion of NPCs in S-phase expressing indicated dSpCas9 and gRNAs at baseline, analyzed by high content imaging, as in . n=7 wells per condition, nine tiled images per well, significance determined by two sample t-test. c) Change in the proportion of NPCs in S-phase in response to 1.5mM LiCl, while expressing indicated dSpCas9 and gRNAs. n=7 wells per condition, significance determined by two sample t-test.

Article Snippet: gRNAs were selected using the GPP sgRNA Designer (Broad Institute) and cloned into CRISPRi (pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-GFP; Addgene # 71237) or CRISPRa (pLV hU6-gRNA (anti-sense) hUbC-VP64-dCas9-VP64-T2A-GFP; Addgene #66707) plasmids, which were kind gifts from the lab of Charles Gersbach.

Techniques: Transduction, Incubation, Expressing, Quantitative RT-PCR, Imaging