Creating CRISPR-Cas9 Knockout Immune Cells using CHIME
Year: 2019
Type: preprint
Abstract: Therapies that target the function of immune cells have signi cant clinical e cacy in diseases such as cancer and autoimmunity.Although functional genomics has accelerated therapeutic target discovery in cancer, its use in primary immune cells is limited because vector delivery is ine cient and can perturb cell states.Here we describe CHIME: CHimeric IMmune Editing, a CRISPR-Cas9 bone marrow delivery system to rapidly evaluate gene function in innate and adaptive immune cells in vivo without ex vivo manipulation of these mature lineages.This approach enables e cient deletion of genes of interest in major immune lineages without altering their development or function.We use this approach to perform an in vivo pooled genetic screen and identify Ptpn2 as a negative regulator of CD8+ T cell-mediated responses to LCMV Clone 13 viral infection.These ndings suggest that this genetic platform can enable rapid target discovery through pooled screening in immune cells in vivo.
Authors: Martin LaFleur, Arlene Sharpe, Thao Nguyen, Matthew Coxe, W. Nicholas Haining
Institution: Harvard University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W4255272201