USP7 CRISPR knock-out in T-ALL cells [Molt4_CRISPR_RNA-seq]

RNAseq analysis of USP7 CRISPR KO in T-ALL cell lines. Genetically modified T-ALL cells were generated using CRISPR-Cas9 technology. Briefly, the sgRNA was designed with at least 3bp of mismatch to any other site in the human genome to mitigate the risk of off-target editing. To generate the modified cells, 400,000 cells were transiently co-transfected with / without 0.5 ug pmaxGFP plasmid (Lonza), with 33 pmol spCas9 protein, 100 pmol chemically modified sgRNA (Synthego), and 100 pmol of blocking ssODN (IDT) via nucleofection (Lonza), using solutions and programs according to the manufacturer's recommended protocol. Five days post-nucleofection, cells were single-cell sorted by FACs into 96-well plates. Cells were clonally expanded and verified the desired modification via targeted deep sequencing Overall design: Partial CRISPR KO of USP7 in Molt4 (n = 2); 100% CRISPR KO of USP7 in Molt4 (n = 2); Wild-type Molt4 (n = 1);