Computational prediction and experimental validation identify functionally conserved lncRNAs from zebrafish to human

Genome-wide CRISPR screens identified lncRNAs that promote cell proliferation in HeLa, Huh7, and MCF7 cells The objectives of this study were to identify lncRNAs that promote cell proliferation in HeLa, Huh7, and MCF7 cells. We performed genome-wide CRISPR knockout screens in HeLa, Huh7, and MCF7 cells using designed pair crRNA library. Initial cell libraries were obtained through lentivirus infection at low MOI (~0.3), followed by collecting GFP-positive cells 72 hours after infection . For each replicate, generally a total of two million GFP-positive cells were collected by FACS . Three replicates of the experimental group were arranged for library screening, and at the same time, another two million GFP-positive cells were collected for genomic DNA extraction as the control group (day 0). During screening, samples containing at least four million cells were collected for genomic DNA extraction at three time points (days 15, 30, and 45) after each cell splitting.