Fig. 7

CRISPRi knockdown of radial glia-enriched lncRNA LOC646329 inhibits proliferation. a RNA-seq alignments at the LOC646329 locus. GW16(+)/(–) replicate one and U87 alignments are number of reads. Radial glia and maturing neurons are merged alignments normalized by number of cells within each cell cluster. sgRNAs targeting the TSS of LOC646329 are indicated. b Expression of radial glia and neuron mRNAs and lncRNAs in U87 glioblastoma cells. c qPCR of LOC646329 following 4 days of CRISPRi knockdown using two sgRNAs targeting the TSS of LOC646329 relative to non-targeting control sgRNA. Biological triplicates (black circles) show 79.0 % repression with sgLOC646329-1 (p = 0.0014; Welch’s t-test) and 62.6 % repression in sgLOC646329-2 (p = 0.0035; Welch’s t-test). Red lines, mean. d Relative growth assays of U87 cells following sgRNA infection. sgRNA⁺ fraction was calculated relative to 5 days post sgRNA infection and normalized to the sgCtrl⁺ fraction at each time point. Biological triplicates show 28.1 % depletion at 20 days with sgLOC646329-1 (p = 0.0073; Welch’s t-test) and 33.5 % depletion with sgLOC646329-2 (p = 0.00048; Welch’s t-test). Error bars, standard deviation of triplicate cultures