Project description:Cells were transfected with plasmids containing AsCas12f variants and sgRNAs for gene-editing. Off-target effect was determined using GUIDE-seq on an Illumina Nextseq platform.
Project description:Mutations in the BRCA2 gene are associated with sporadic and familial cancer, cause genomic instability, and sensitize cancer cells to PARP inhibition. We have developed a human primary cell system to annotate variants in BRCA2 and to test the variants’ sensitivities to PARP inhibition. We engineered locally haploid human pluripotent stem cells (loHAPs) that contain a localized deletion encompassing one copy of BRCA2. Next, we characterized essential regions of the BRCA2 gene to identify permissive and loss-of-function mutations in hPSCs and differentiated fibroblasts, using CRISPR-Cas9 editing of the functional allele. Additionally, we used gene editing to directly test the function of individual amino acids, including clinical variants of uncertain significance in BRCA2, and identify alleles that are sensitive PARP inhibitors, a standard of care in BRCA2-deficient cancers. Collectively, our analysis demonstrates that loHAPs can facilitate detailed structure-function analysis of genes and the rapid functional evaluation of clinically-observed mutations.
2023-12-31 | GSE233683 | GEO
Project description:Engineered APOBEC3A deaminase variants for highly accurate and efficient cytosine base editing
| PRJNA938580 | ENA
Project description:Engineered APOBEC3A deaminase variants for highly accurate and efficient cytosine base editing
| PRJNA1032216 | ENA
Project description:Engineered APOBEC3A deaminase variants for highly accurate and efficient cytosine base editing
| PRJNA938578 | ENA
Project description:Engineered APOBEC3A deaminase variants for highly accurate and efficient cytosine base editing
| PRJNA938162 | ENA
Project description:Engineered APOBEC3A deaminase variants for highly accurate and efficient cytosine base editing
Project description:To compare the impact of TP53 mutant variants in exon-wide mutant cell libraries, comprehensive mutome libraries of TP53 affecting the Exons 5, 6, 7 and 8 were generated systematically by CRISP/Cas9 editing in HCT116 colorectal carcinoma cells.
