LuCaP 23.1 ectopically expressing HNF1A, rep4
PDX tumors were passaged in NOD scid gamma (NSG) mice at the NCI and propagated in vitro as 3-dimensional organoid cultures. PDXs were enzymatically degraded using the MACS human Tumor Dissociation Kit. LuCaP 23.1 cells were transduced with generated lentivirus via centrifugation and LentiBOOST. RNA was extracted from first generation organoids using RNeasy kit according to manufacturer's instructions, and profiled for quality using a Bioanalyzer. Poly-A–enriched and Illumina–barcoded libraries were prepared using the TruSeq kit. Libraries were sequenced on a NextSeq 500 to a depth of 75 million base pairs (150 bp paired end) at the NCI Sequencing Facility.