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Synthetic type III-E CRISPR-Cas effectors for programmable RNA-targeting
Open accessPreprint — published as 10.1016/j.jmb.2025.169566 · Paper page
Abstract
The recent discovery of the type III-E CRISPR-Cas effector class has reshaped our fundamental understanding of CRISPR-Cas evolution and classification. Type III-E effectors are composed of several Cas7-like domains and a single Cas11-like domain naturally fused together to create a single polypeptide capable of programmably targeting and degrading RNA. Here we identify a novel composition of a type III-E-like effector composed of Cas7-like and a Cas1-like domain, that can be engineered into an active chimeric RNA-targeting Cas effector and presents a new function of Cas1 in RNA-targeting. Furthermore, we demonstrate a unique modularity of type III-E effectors by methodically substituting domains between orthogonal type III-E proteins to engineer compact synthetic Cas effectors. We refine our methods to generate several compact effectors for programmable RNA-targeting in mammalian cells. Cas7-S represents a new understanding of type III-E architecture and modularity, and provides a platform for engineering genome engineering technologies from the blueprint of nature.
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Metadata
- Authors
- Brogan, D. J., Benetta, E. D., Wang, T., Lin, C. P., Chen, F., Li, H., Lin, C., Komives, E. A., & Akbari, O. S.
- Affiliations
- University of California San Diego (US)
- Corresponding author
- Komives, Elizabeth A.
- Akbari, Omar S.
- ORCID
- Venue
- Journal of Molecular Biology
- Publisher
- openRxiv
- Published
- 2024-02-24
- Volume
- 438
- Issue
- 2
- Pages
- 169566-169566
- ISSN
- 0022-2836 (ISSN-L)
- PMC
- PMC13404073
- OpenAlex
- W4416746565
- Cited by
- 4
- References
- 1
- Citations by year
- 2025: 2 · 2024: 2
- FWCI
- 0.60
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- cc-by-nc-nd
- Locations
- Journal of Molecular Biology (publishedVersion, cc-by-nc-nd)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed (publishedVersion)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- Funders
- National Institutes of Health (1dp2ai152071-01)
- National Institute of General Medical Sciences (R01GM132825)
- National Institute of Allergy and Infectious Diseases (R21AI149161, RO1AI1900001)
- Division of Microbiology and Infectious Diseases, National Institute of Allergy and Infectious Diseases (R21RAI149161A, R01GM132825-04, DP2AI152071)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- domain swapping, RNA degradation
- MeSH terms
- Protein Domains, Gene Editing, RNA, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Cas Systems, CRISPR-Associated Proteins
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