Genome-wide CRISPR interference screen identifies long non-coding RNA loci required for differentiation and pluripotency
收藏干细胞与再生医学数据中心2022-02-20 更新2024-03-06 收录
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Systematic identification of factors that regulate human embryonic stem cell (hESC) differentiation can provide valuable insights into early development and disease. However, effective modulation of gene expression during differentiation has proven technically challenging. Here, we report the first described hESC lines to repress (CRISPRi) or activate (CRISPRa) transcription during differentiation into all three germ layers. We subsequently performed an unbiased, genome-wide CRISPRi screen targeting long non-coding RNAs during endoderm differentiation. From our screen, we identified known factors, as well as novel lncRNAs required for differentiation. Surprisingly, lncRNA expression was not predictive of enrichment scores, supporting the necessity for functional screening instead of relying on gene expression analysis. Further downstream analysis of two hits, RP11-222K16.2 and FOXD3-AS1, revealed two lncRNAs essential for differentiation and pluripotency, respectively. Taken together, the cell lines and screen methodology described herein can be adapted to discover novel regulators of differentiation into any lineage.
提供机构：
Harvard University
创建时间：
2022-02-20
