Distinct immunomodulatory pathways mediate radiotherapy response in malignant peripheral nerve sheath tumors [sgRNA library]

We performed a genome-wide CRISPRi screen to identify potential contributors to radioresistance in the JH02.2 malignant peripheral nerve sheath tumor (MPNST) cell line. Overall design: We transduced a barcoded lentiviral sgRNA library into a JH-2-002 human MPNST cell line expressing dCas9-Zim3, with virus titrated to approximate 1 sgRNA per cell. Cells were sorted for library expression using FACS, and a T0 control was collected at Day 0. Two experimental groups were designed, with one receiving 2 daily fractions of 2 Gy and the other being untreated, receiving no radiation. Genomic DNA (gDNA) from all cells was harvested at Day 10 and sgRNA barcodes were amplified and sequenced.