RBP-CRISPR data of "In vivo CRISPR screen identifies SNRPC as a regulator for facilitating triple-negative breast cancer progression"

In this study, we constructed a CRISPR/Cas9 library targeting 1114 RBPs for systematic screening. The DNA oligonucleotide library contained 8667 sgRNA sequences (7667 sgRNAs targeting 1114 genes+1000 control sgRNAs). Full-length oligonucleotides were amplified by PCR and cloned by Gibson ligation reaction for the assembly into the lentiGuide-Puro vector. Then, the ligation products were transformed into the electrocompetent cells. The viral RBP library was produced in HEK-293T cells. DCIS and CA1a cells stably expressing lentiCas9-Blast were infected with the virus mixture at a low multiplicity of infection (MOI=0.3). The infected cells were then selected with 1-2ug/mL puromycin for 7 days. We divided mixed cloned cells into 6 parts: 3 parts represented baseline cells on day0, whose genomic DNA was immediately extracted; 3 parts were inoculated subcutaneously into the mammary fat pad of six-week-old female NOD-SCID mice to generate transplanted tumor models. After 4 weeks of growth, the subcutaneous tumors were excised and their DNA was extracted.