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Molecular and Cellular Biology, Genetics

Abstract 403: CRISPR/Cas9 genome-wide sgRNA libraries for loss-of-function and gain-of-function genetic screens

Donato Tedesco, Paul Diehl, Mikhail Makhanov, Sylvain Baron and Alex Chenchik
Donato Tedesco
Cellecta, Inc., Mountain View, CA.
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Paul Diehl
Cellecta, Inc., Mountain View, CA.
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Mikhail Makhanov
Cellecta, Inc., Mountain View, CA.
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Sylvain Baron
Cellecta, Inc., Mountain View, CA.
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Alex Chenchik
Cellecta, Inc., Mountain View, CA.
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DOI: 10.1158/1538-7445.AM2017-403 Published July 2017
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Proceedings: AACR Annual Meeting 2017; April 1-5, 2017; Washington, DC

Abstract

Genome-wide loss-of-function screening is a fundamental method to identify genes responsible for driving biological processes. Complex pooled lentiviral-based libraries expressing large numbers of genetic disruptors, such as shRNAs (RNAi) or sgRNAs (CRISPR), make large-scale cell screening practical. While RNAi-based approaches have proven to be an effective strategy for these screens, recent work has showed CRISPR-based technologies offer not only an effective alternative, but distinct advantage. We will present unpublished data from genetic screens in human cancer cell lines using CRISPR-Knockout (KO), CRISPRi and CRISPRa technologies, as well as RNAi. Although loss-of-function shRNA and sgRNA pooled library screens are similar in concept, the gene loss of function is achieved by different mechanisms (mRNA degradation with RNAi, full gene disruption with CRISPR-KO, transcriptional inhibition with CRISPRi), so some divergences are expected and indeed observed when comparing results obtained using one method versus the other. Furthermore, contrary to RNAi, CRISPR technology can be modified to activate gene expression (CRISPRa), thus enabling the use of genome-wide gain-of-function screening in gene function studies.

Citation Format: Donato Tedesco, Paul Diehl, Mikhail Makhanov, Sylvain Baron, Alex Chenchik. CRISPR/Cas9 genome-wide sgRNA libraries for loss-of-function and gain-of-function genetic screens [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 403. doi:10.1158/1538-7445.AM2017-403

  • ©2017 American Association for Cancer Research.
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Cancer Research: 77 (13 Supplement)
July 2017
Volume 77, Issue 13 Supplement
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Abstract 403: CRISPR/Cas9 genome-wide sgRNA libraries for loss-of-function and gain-of-function genetic screens
Donato Tedesco, Paul Diehl, Mikhail Makhanov, Sylvain Baron and Alex Chenchik
Cancer Res July 1 2017 (77) (13 Supplement) 403; DOI: 10.1158/1538-7445.AM2017-403

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Abstract 403: CRISPR/Cas9 genome-wide sgRNA libraries for loss-of-function and gain-of-function genetic screens
Donato Tedesco, Paul Diehl, Mikhail Makhanov, Sylvain Baron and Alex Chenchik
Cancer Res July 1 2017 (77) (13 Supplement) 403; DOI: 10.1158/1538-7445.AM2017-403
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Cancer Research Online ISSN: 1538-7445
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