CRISPR-based functional genomics in human dendritic cells
Name
elife-65856-v2.pdf
Description
Published version
Size
1.18 MB
Format
Unknown
Checksum (MD5)
46b50d354b7fed02a2363b9dc0c5fbda
Author(s)
Jost, Marco
Jacobson, Amy N
Hussmann, Jeffrey A
Cirolia, Giana
Fischbach, Michael A
Weissman, Jonathan S
Date Issued
April 2021
Journal
eLife
Publisher
eLife Sciences Publications, Ltd
Version
Final published version
Abstract
Dendritic cells (DCs) regulate processes ranging from antitumor and antiviral immunity to host-microbe communication at mucosal surfaces. It remains difficult, however, to genetically manipulate human DCs, limiting our ability to probe how DCs elicit specific immune responses. Here, we develop a CRISPR-Cas9 genome editing method for human monocyte-derived DCs (moDCs) that mediates knockouts with a median efficiency of >94% across >300 genes. Using this method, we perform genetic screens in moDCs, identifying mechanisms by which DCs tune responses to lipopolysaccharides from the human microbiome. In addition, we reveal donor-specific responses to lipopolysaccharides, underscoring the importance of assessing immune phenotypes in donor-derived cells, and identify candidate genes that control this specificity, highlighting the potential of our method to pinpoint determinants of inter-individual variation in immunity. Our work sets the stage for a systematic dissection of the immune signaling at the host-microbiome interface and for targeted engineering of DCs for neoantigen vaccination.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Whitehead Institute for Biomedical Research
Terms of Use
Creative Commons Attribution 4.0 International license
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.7554/elife.65856
