CRISPR screen to determine host factors
Discover host factors critical for SARS-CoV-2 pathogenesis and identify novel therapeutic targets
CRISPR screens in the field of virology are ideally suited for analyzing virus-host interactions; determining host factors critical for virus entry, replication, and spread; and identifying and prioritizing novel drug targets. Performing a genome-scale CRISPR/Cas9 knockout screen with an sgRNA library can help identify important host factors responsible for disease response.
The Guide-it CRISPR Genome-Wide sgRNA Library System is a pooled lentiviral sgRNA library for performing phenotypic knockout screens in human cell lines. The system is based on the Brunello library (Doench et al. 2016; Doench et al. 2018), and contains over 76,000 guide RNAs (sgRNAs) targeting about 19,000 genes, with four highly active guides per gene, as well as nontargeting sgRNA controls. The amplified, validated library is provided in a ready-to-transfect lyophilized format enabling easy production of high-titer lentivirus and setup of genome-wide CRISPR screens (Figure 1).
Following screening, next-generation sequencing (NGS) analysis can be performed to identify candidate genes of interest using the Guide-it CRISPR Genome-Wide sgRNA Library NGS Analysis Kit, which includes the necessary primers and reagents to prepare Illumina-sequencing-ready NGS libraries from cell populations transduced with our genome-wide sgRNA library.
Quality control to ensure sgRNA representation
Our library has been carefully manufactured to ensure proper sgRNA representation. We have measured sgRNA representation at every step in the process: library synthesis, amplification, viral production, and target cell transduction (Figure 2).
Examples of studies using genome-wide CRISPR screens for virology
Flint, M. A genome-wide CRISPR screen identifies N-acetylglucosamine-1-phosphate transferase as a potential antiviral target for Ebola virus. Nat. Comm. 285, (2019).
Gebre, M. et al. CRISPR-Cas9 Genetic Analysis of Virus-Host Interactions. Viruses 10, 55 (2018).
Han, J. Genome-wide CRISPR/Cas9 Screen Identifies Host Factors Essential for Influenza Virus Replication. Cell Rep. 23, 596–607 (2018).
Li, B. et al. Genome-wide CRISPR screen identifies host dependency factors for influenza A virus infection. Nat. Comm. 11, 164 (2020).
Marceau, C. D. Genetic dissection of Flaviviridae host factors through genome-scale CRISPR screens. Nature 535, 159–163 (2016).
Park, R. J. A genome-wide CRISPR screen identifies a restricted set of HIV host dependency factors. Nat. Genet. 49, 193–203 (2017).
Puschnik, A. S. et al. A CRISPR toolbox to study virus-host interactions. Nat. Rev. Microbiol. 15, 351–364 (2017).
Doench, J. G. et al. Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9. Nat. Biotechnol. 34, 184–191 (2016).
Doench, J. G. et al. Am I ready for CRISPR? A user's guide to genetic screens. Nat. Rev. Genet. 19, 67–80 (2018).
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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