A CRISPR-Cas9 delivery system for in vivo screening of genes in the immune system

Nat Commun. 2019 Apr 10;10(1):1668. doi: 10.1038/s41467-019-09656-2.

Abstract

Therapies that target the function of immune cells have significant clinical efficacy 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 inefficient 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 efficient 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 findings indicate that this genetic platform can enable rapid target discovery through pooled screening in immune cells in vivo.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptive Immunity / genetics*
  • Animals
  • Bone Marrow Transplantation
  • CD8-Positive T-Lymphocytes / immunology
  • CRISPR-Cas Systems / genetics*
  • Cell Line, Tumor
  • Cell Lineage / genetics
  • Cell Lineage / immunology
  • Chlorocebus aethiops
  • Disease Models, Animal
  • Feasibility Studies
  • Female
  • Gene Transfer Techniques*
  • Genetic Testing / methods*
  • Genetic Vectors / genetics
  • Genomics / methods
  • HEK293 Cells
  • Humans
  • Immunity, Innate / genetics*
  • Lymphocytic Choriomeningitis / genetics
  • Lymphocytic Choriomeningitis / immunology
  • Lymphocytic Choriomeningitis / virology
  • Lymphocytic choriomeningitis virus / immunology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Protein Tyrosine Phosphatase, Non-Receptor Type 2 / genetics
  • Protein Tyrosine Phosphatase, Non-Receptor Type 2 / immunology
  • RNA, Guide / genetics
  • Transplantation Chimera
  • Vero Cells

Substances

  • RNA, Guide
  • PTPN2 protein, human
  • Protein Tyrosine Phosphatase, Non-Receptor Type 2