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Optimized RNA-targeting CRISPR/Cas13d technology outperforms shRNA in identifying functional circRNAs

Zhang, Yang
; 
Nguyen, Tuan M.
; 
Zhang, Xiao-Ou
; 
Wang, Limei
; 
Phan, Tin
; 
Clohessy, John G.
; 
Pandolfi, Pier Paolo
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Authors
Zhang, Yang
Nguyen, Tuan M.
Zhang, Xiao-Ou
Wang, Limei
Phan, Tin
Clohessy, John G.
Pandolfi, Pier Paolo
Student Authors
Faculty Advisor
Academic Program
UMass Chan Affiliations
Program in Bioinformatics and Integrative Biology
Document Type
Journal Article
Publication Date
2021-01-21
Keywords
CRISPR/Cas13d system
 High-throughput screening
 circRNAs
 Bioinformatics
 Genetics and Genomics
 Nucleic Acids, Nucleotides, and Nucleosides
Subject Area
Collections
UMass Chan Faculty and Researcher Publications
Genomics and Computational Biology Publications
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Abstract

Short hairpin RNAs (shRNAs) are used to deplete circRNAs by targeting back-splicing junction (BSJ) sites. However, frequent discrepancies exist between shRNA-mediated circRNA knockdown and the corresponding biological effect, querying their robustness. By leveraging CRISPR/Cas13d tool and optimizing the strategy for designing single-guide RNAs against circRNA BSJ sites, we markedly enhance specificity of circRNA silencing. This specificity is validated in parallel screenings by shRNA and CRISPR/Cas13d libraries. Using a CRISPR/Cas13d screening library targeting > 2500 human hepatocellular carcinoma-related circRNAs, we subsequently identify a subset of sorafenib-resistant circRNAs. Thus, CRISPR/Cas13d represents an effective approach for high-throughput study of functional circRNAs.

Source

Zhang Y, Nguyen TM, Zhang XO, Wang L, Phan T, Clohessy JG, Pandolfi PP. Optimized RNA-targeting CRISPR/Cas13d technology outperforms shRNA in identifying functional circRNAs. Genome Biol. 2021 Jan 21;22(1):41. doi: 10.1186/s13059-021-02263-9. PMID: 33478577; PMCID: PMC7818937. Link to article on publisher's site

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DOI
10.1186/s13059-021-02263-9
Permanent Link to this Item
https://hdl.handle.net/20.500.14038/41744
PubMed ID
33478577
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© The Author(s). 2021 Open Access: This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
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