Journal
NATURE COMMUNICATIONS
Volume 10, Issue -, Pages -Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/s41467-018-08158-x
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Funding
- Stanford School of Medicine Postdoctoral Fellowship
- ALS Association Milton Safenowitz Fellowship
- Burroughs Wellcome Fund Postdoctoral Enrichment Program
- National Science Foundation Graduate Research Fellowship Program
- Stanford-NIST Joint Initiative in Molecular Biology training grant
- EDGE-STEM fellowship from the Stanford Vice Provost of Graduate Education
- Pew Charitable Trusts
- Alfred P. Sloan Foundation
- Li Ka Shing Foundation
- Stanford University
- Stanford Bioengineering department
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Repurposed CRISPR-Cas molecules provide a useful tool set for broad applications of genomic editing and regulation of gene expression in prokaryotes and eukaryotes. Recent discovery of phage-derived proteins, anti-CRISPRs, which serve to abrogate natural CRISPR anti-phage activity, potentially expands the ability to build synthetic CRISPR-mediated circuits. Here, we characterize a panel of anti-CRISPR molecules for expanded applications to counteract CRISPR-mediated gene activation and repression of reporter and endogenous genes in various cell types. We demonstrate that cells pre-engineered with anti-CRISPR molecules become resistant to gene editing, thus providing a means to generate write-protected cells that prevent future gene editing. We further show that anti-CRISPRs can be used to control CRISPR-based gene regulation circuits, including implementation of a pulse generator circuit in mammalian cells. Our work suggests that anti-CRISPR proteins should serve as widely applicable tools for synthetic systems regulating the behavior of eukaryotic cells.
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