Saturation Genome Editing of BAP1

ABSTRACT: Saturation Genome Editing of BRCA1 Associated Protein 1 (BAP1). This study aims to functionally characterize saturating-levels of mutation at the BAP1 locus. We will target the Coding Sequence (CDS), splice-site containing intron and 3'UTR of the transcript: ENST00000460680.6. CRISPR-Cas9 and variant harbouring dsDNA repair templates will be co-transfected into HAP1 cells (Lig4-& clonal Cas9 expressing, sorted for 1n ploidy) to edit populations of cells. Duplicate libraries will be used to target most regions (that is the same oligonucleotide library composition for a region, but with a different sgRNA used for the CRISPR-Cas9 component). Targeted regions will then be sampled with gDNA extracted from cells at day 4, 7, 11, 14 & 21 after transfection. Amplicons are then generated for each o

ORGANISM(S): Homo sapiens (human)

SUBMITTER:  

PROVIDER: S-BSST1222 | biostudies-other |

REPOSITORIES: biostudies-other