Perturbation of long non-coding RNA therapeutic targets in glioma

We used CRISPR interference (CRISPRi) to screen 5689 lncRNA loci in human glioblastoma (GBM) cells, identifying 467 hits that modified cell growth in the presence of clinically relevant doses of fractionated radiation. 33 of these lncRNA hits sensitized cells to radiation, and based on their expression in adult and pediatric glioma, nine of these hits were prioritized as lncRNA Glioma Radiation Sensitizers (lncGRS). Knockdown lncGRS-1, a primate-conserved, nuclear-enriched lncRNA, inhibited the growth and proliferation of primary adult and pediatric glioma cells, but not the viability of normal brain cells. RNA-seq was performed following lncGRS-1 knockdown in 3 different cell lines under normal conditions. Long-read single molecule native RNA sequencing was also performed on untreated U87 cell cultures using the Oxford Nanopore PromethION.