Abstract
CRISPR-mediated interference (CRISPRi), a robust and specific system for programmably repressing transcription, provides a versatile tool for systematically characterizing the function of long noncoding RNAs (lncRNAs). When used with highly parallel, lentiviral pooled screening approaches, CRISPRi enables the targeted knockdown of tens of thousands of lncRNA-expressing loci in a single screen. Here we describe the use of CRISPRi to target lncRNA loci in a pooled screen, using cell growth and proliferation as an example of a phenotypic readout. Considerations for custom lncRNA-targeting libraries, alternative phenotypic readouts, and orthogonal validation approaches are also discussed.
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Liu, S.J., Horlbeck, M.A., Weissman, J.S., Lim, D.A. (2021). Genome-Scale Perturbation of Long Noncoding RNA Expression Using CRISPR Interference. In: Cao, H. (eds) Functional Analysis of Long Non-Coding RNAs. Methods in Molecular Biology, vol 2254. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1158-6_20
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DOI: https://doi.org/10.1007/978-1-0716-1158-6_20
Publisher Name: Humana, New York, NY
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