PT  - JOURNAL ARTICLE
AU  - Soyeong Jun
AU  - Hyeonseob Lim
AU  - Honggu Chun
AU  - Ji Hyun Lee
AU  - Duhee Bang
TI  - Single-cell analysis of a mutant library generated using CRISPR-guided deaminase
AID  - 10.1101/610725
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 610725
4099  - http://biorxiv.org/content/early/2020/01/21/610725.short
4100  - http://biorxiv.org/content/early/2020/01/21/610725.full
AB  - CRISPR-based screening methods using single-cell RNA sequencing (scRNA-seq) technology enable comprehensive profiling of gene perturbations from knock-out mutations. However, evaluating substitution mutations using scRNA-seq is currently limited. We combined CRISPR RNA-guided deaminase and scRNA-seq technology to develop a platform for introducing mutations in multiple genes and assessing the mutation-associated signatures. Using this platform, we generated a library consisting of 420 sgRNAs, performed sgRNA tracking analysis, and assessed the effect size of the response to vemurafenib in the melanoma cell line, which has been well-studied via knockout-based drop-out screens. However, a substitution mutation library screen has not been applied and transcriptional information for mechanisms of action was not assessed. Our platform permits discrimination of several candidate mutations that function differently from other mutations by integrating sgRNA candidates and gene expression readout. We anticipate that our platform will enable high-throughput analyses of the mechanisms related to a variety of biological events.