CRISPR-CasX is an RNA-dominated enzyme active for human genome editing

By Jun-Jie Liu, N. Orlova, Benjamin L. Oakes, E. Ma, Hannah B. Spinner, Katherine L. M. Baney, Jonathan Chuck, D. Tan, G. Knott, Lucas B. Harrington and 8 more. 2019

360 citations recorded by the source corpus.

The RNA-guided CRISPR-associated (Cas) proteins Cas9 and Cas12a provide adaptive immunity against invading nucleic acids, and function as powerful tools for genome editing in a wide range of organisms. Here we reveal the underlying mechanisms of a third, fundamentally distinct RNA-guided genome-editing platform named CRISPR–CasX, which uses unique structures for programmable double-stranded DNA binding and cleavage. Biochemical and in vivo data demonstrate that CasX is active for Escherichia coli and human genome modification. Eight cryo-electron microscopy structures of CasX in different states of assembly with its guide RNA and double-stranded DNA substrates reveal an extensive RNA scaffold and a domain required for DNA unwinding. These data demonstrate how CasX activity arose through convergent evolution to establish an enzyme family that is functionally separate from both Cas9 and Cas12a. CRISPR–CasX represents a distinct RNA-guided platform that is functionally separate from Cas9 and Cas12a and is active for bacterial and human genome modification.

Subfields: Molecular Biology

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In Molecular Biology

1. Michael Jendrusch
2. Yan Duan
3. Yun S. Song
4. Pieter Abbeel
5. John F. Canny
6. Neil Thomas
7. Xi Chen
8. Roshan Rao

Citations in the corpus

• Evaluating Protein Transfer Learning with TAPE (2019, background): “(Impact) Detection of remote homologs is of great interest in microbiology and medicine; e.g., for detection of emerging antibiotic resistant genes [41] and discovery of new CAS enzymes [42].”
• Leveraging implicit knowledge in neural networks for functional dissection and engineering of proteins (2019, methodology): “First, we analysed DeeProtein’s sensitivity for Cas9 nuclease activity in the RuvC catalytic domain, which is conserved in Cas orthologues, including the recently discovered CasX class 36 .”