A Three-Way Combinatorial CRISPR Screen for Analyzing Interactions among Druggable Targets

Peng Zhou; Becky K C Chan; Yuk Kei Wan; Chaya T L Yuen; Gigi C G Choi; Xinran Li; Cindy S W Tong; Sophia S W Zhong; Jieran Sun; Yufan Bao; Silvia Y L Mak; Maggie Z Y Chow; Jien Vei Khaw; Suet Yi Leung; Zongli Zheng; Lydia W T Cheung; Kaeling Tan; Koon Ho Wong; H Y Edwin Chan; Alan S L Wong

doi:10.1016/j.celrep.2020.108020

A Three-Way Combinatorial CRISPR Screen for Analyzing Interactions among Druggable Targets

Cell Rep. 2020 Aug 11;32(6):108020. doi: 10.1016/j.celrep.2020.108020.

Authors

Peng Zhou¹, Becky K C Chan¹, Yuk Kei Wan¹, Chaya T L Yuen¹, Gigi C G Choi¹, Xinran Li², Cindy S W Tong¹, Sophia S W Zhong¹, Jieran Sun¹, Yufan Bao³, Silvia Y L Mak⁴, Maggie Z Y Chow⁴, Jien Vei Khaw¹, Suet Yi Leung⁵, Zongli Zheng⁶, Lydia W T Cheung², Kaeling Tan⁷, Koon Ho Wong⁸, H Y Edwin Chan⁹, Alan S L Wong¹⁰

Affiliations

¹ Laboratory of Combinatorial Genetics and Synthetic Biology, School of Biomedical Sciences, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
² School of Biomedical Sciences, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
³ Ming Wai Lau Centre for Reparative Medicine, Karolinska Institutet, Hong Kong SAR, China; Department of Biomedical Sciences, City University of Hong Kong, Hong Kong SAR, China.
⁴ Ming Wai Lau Centre for Reparative Medicine, Karolinska Institutet, Hong Kong SAR, China.
⁵ Department of Pathology, The University of Hong Kong, Pokfulam, Hong Kong SAR, China; Centre for PanorOmic Sciences, The University of Hong Kong, Pokfulam, Hong Kong SAR, China; The Jockey Club Centre for Clinical Innovation and Discovery, LKS Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
⁶ Ming Wai Lau Centre for Reparative Medicine, Karolinska Institutet, Hong Kong SAR, China; Department of Biomedical Sciences, City University of Hong Kong, Hong Kong SAR, China; Biotechnology and Health Centre, City University of Hong Kong Shenzhen Research Institute, Shenzhen, China.
⁷ Faculty of Health Sciences, University of Macau, Macau SAR, China; Genomics, Bioinformatics and Single Cell Analysis Core, Faculty of Health Sciences, University of Macau, Avenida da Universidade, Taipa, Macau SAR, China.
⁸ Faculty of Health Sciences, University of Macau, Macau SAR, China; Institute of Translational Medicine, University of Macau, Avenida da Universidade, Taipa, Macau SAR, China.
⁹ Laboratory of Drosophila Research, School of Life Sciences, The Chinese University of Hong Kong, Hong Kong SAR, China; Gerald Choa Neuroscience Centre, The Chinese University of Hong Kong, Hong Kong SAR, China.
¹⁰ Laboratory of Combinatorial Genetics and Synthetic Biology, School of Biomedical Sciences, The University of Hong Kong, Pokfulam, Hong Kong SAR, China; Department of Electrical and Electronic Engineering, The University of Hong Kong, Pokfulam, Hong Kong SAR, China. Electronic address: aslw@hku.hk.

PMID: 32783942
DOI: 10.1016/j.celrep.2020.108020

Abstract

We present a CRISPR-based multi-gene knockout screening system and toolkits for extensible assembly of barcoded high-order combinatorial guide RNA libraries en masse. We apply this system for systematically identifying not only pairwise but also three-way synergistic therapeutic target combinations and successfully validate double- and triple-combination regimens for suppression of cancer cell growth and protection against Parkinson's disease-associated toxicity. This system overcomes the practical challenges of experimenting on a large number of high-order genetic and drug combinations and can be applied to uncover the rare synergistic interactions between druggable targets.

Keywords: CRISPR; CombiGEM; Parkinson's disease; cancer; combination therapy; combinatorial genetics; drug interaction; genetic interaction; high-throughput screening; neurodegeneration.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antineoplastic Agents / pharmacology
CRISPR-Cas Systems*
Drosophila melanogaster
Drug Combinations*
Drug Delivery Systems / methods*
Gene Knockout Techniques
HEK293 Cells
High-Throughput Screening Assays / methods*
Humans
Mice
Neoplasms / drug therapy
Parkinson Disease / drug therapy
RNA, Guide, CRISPR-Cas Systems

Substances

Antineoplastic Agents
Drug Combinations
RNA, Guide, CRISPR-Cas Systems