New Strategies for Forward Genetic Screening and Single Cell Analysis with CRISPRi Transcriptional Repressors
Year: 2020
Type: article
Abstract: The power of CRISPR-Cas9 for genome engineering has continued to grow in recent years. The ability to perform large-scale, whole genome loss-of-function screens has allowed for new breakthroughs identifying gene pathways in drug resistance and disease. CRISPRi (inhibition) allows for targeted suppression of gene function by delivering transcriptional repressors to a specific target sequence using modified dCas9/gRNA complexes. Gene knockdown is complementary to both existing loss-of-function and gain-of-function technologies; and has several distinct advantages, including efficient targeting of non-coding genes. We have developed whole genome CRISPRi libraries for high throughput screening, divided into functionally relevant subpools for a complementary or alternative screening approach. Furthermore, and in partnership with 10X Genomics, we have developed compatible CRISPR vectors for single cell genomic exploration. These CRISPR guide vectors contain capture sequences that allow efficient feature barcoding of individual cells for transcriptomics across pools of guides. This provides key advantages in functional genomic analysis of top screen hits.
Source: PubMed Central
Authors: Ravanelli, Andrew, Meer, Elliott, Terry, Jessica, Ji, Qingzhou, Ashworth, Roxann, Chittur, Sridar
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7021158486