A genome-wide knockdown screen for host factors that participate in CRISPR adaptation reveals novel factors required for spacer acquisition
Board Location: #164
Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology
Session: 2
Yumie Lee - University of California, Merced
Co-Author(s): Santiago C. Lopez, Graduate Program in Bioengineering, University of California, San Francisco and Berkeley, CA, USA Gladstone Institute of Data Science and Biotechnology; Karen Zhang, Graduate Program in Bioengineering, University of California, San Francisco and Berkeley, CA, USA Gladstone Institute of Data Science and Biotechnology; Seth L. Shipman, Graduate Program in Bioengineering, University of California, San Francisco and Berkeley, CA, USA Gladstone Institute of Data Science and Biotechnology
The CRISPR-Cas-system works as an adaptive immune system that defends prokaryotes against foreign genetic material. Adaptation, the initial stage, integrates short fragments of foreign DNA called spacers into the host’s CRISPR array. In Escherichia coli, the Cas1-Cas2 complex interacts with the integration host factor (IHF) to enable spacer addition into the CRISPR array1. However, there may be other essential factors that participate in the adaptation process. To obtain a more complete picture of the components that participate in CRISPR adaptation, a genome-wide CRISPR knockdown screen was explored. Our screen uncovered novel proteins necessary for the acquisition of new spacers. The results of our project direct us toward a better understanding of CRISPR adaptation.
Funder Acknowledgement(s): Gladstone Institutes UCSF Program for Breakthrough Biomedical Research The PEW - charitable trusts Simons Foundation Autism Research Initiative NSFNIH - New Innovator award Chan Zuckerberg Biohub
Faculty Advisor: Seth Shipman, seth.shipman@gladstone.ucsf.edu
Role: I ran a knockdown screen, and the PCR and gel electrophoresis validations using knockout strains.