title: A CRISPR-Based Toolbox for Studying T Cell Signal Transduction.
abstract: CRISPR/Cas9 system is a powerful technology to perform genome editing in a variety of cell types. To facilitate the application of Cas9 in mapping T cell signaling pathways, we generated a toolbox for large-scale genetic screens in human Jurkat T cells. The toolbox has three different Jurkat cell lines expressing distinct Cas9 variants, including wild-type Cas9, dCas9-KRAB, and sunCas9. We demonstrated that the toolbox allows us to rapidly disrupt endogenous gene expression at the DNA level and to efficiently repress or activate gene expression at the transcriptional level. The toolbox, in combination with multiple currently existing genome-wide sgRNA libraries, will be useful to systematically investigate T cell signal transduction using both loss-of-function and gain-of-function genetic screens.
authors.affiliation: School of Life Science and Technology, ShanghaiTech University, Shanghai 200031, China.
authors.forename: Shen
authors.initials: S
authors.lastname: Chi
authors.affiliation: Division of Rheumatology, Department of Medicine, Rosalind Russell Medical Research Center for Arthritis, University of California, San Francisco, CA 94143, USA; Howard Hughes Medical Institute, University of California, San Francisco, CA 94143, USA.
authors.forename: Arthur
authors.initials: A
authors.lastname: Weiss
authors.affiliation: School of Life Science and Technology, ShanghaiTech University, Shanghai 200031, China.
authors.forename: Haopeng
authors.initials: H
authors.lastname: Wang
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics*; D019295:Computational Biology / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D019169:Jurkat Cells; D015398:Signal Transduction / Q000235:genetics* / Q000502:physiology*; D012984:Software; D013601:T-Lymphocytes / Q000502:physiology*