Compact RNA editors with small Cas13 proteins

Soumya Kannan; Han Altae-Tran; Xin Jin; Victoria J Madigan; Rachel Oshiro; Kira S Makarova; Eugene V Koonin; Feng Zhang

doi:10.1038/s41587-021-01030-2

Compact RNA editors with small Cas13 proteins

Nat Biotechnol. 2022 Feb;40(2):194-197. doi: 10.1038/s41587-021-01030-2. Epub 2021 Aug 30.

Authors

Soumya Kannan^#^{1

2

3

4

5}, Han Altae-Tran^#^{1

2

3

4

5}, Xin Jin^{1

2

3

4

5

6

7}, Victoria J Madigan^{1

2

3

4

5}, Rachel Oshiro^{1

2

3

4

5}, Kira S Makarova⁸, Eugene V Koonin⁸, Feng Zhang^{9

10

11

12

13}

Affiliations

¹ Howard Hughes Medical Institute, Cambridge, MA, USA.
² Broad Institute of MIT and Harvard, Cambridge, MA, USA.
³ McGovern Institute for Brain Research at MIT, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁴ Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁵ Department of Brain and Cognitive Science, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁶ Society of Fellows, Harvard University, Cambridge, MA, USA.
⁷ Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA.
⁸ National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, MD, USA.
⁹ Howard Hughes Medical Institute, Cambridge, MA, USA. [email protected].
¹⁰ Broad Institute of MIT and Harvard, Cambridge, MA, USA. [email protected].
¹¹ McGovern Institute for Brain Research at MIT, Massachusetts Institute of Technology, Cambridge, MA, USA. [email protected].
¹² Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. [email protected].
¹³ Department of Brain and Cognitive Science, Massachusetts Institute of Technology, Cambridge, MA, USA. [email protected].

^# Contributed equally.

Abstract

CRISPR-Cas13 systems have been developed for precise RNA editing, and can potentially be used therapeutically when temporary changes are desirable or when DNA editing is challenging. We have identified and characterized an ultrasmall family of Cas13b proteins-Cas13bt-that can mediate mammalian transcript knockdown. We have engineered compact variants of REPAIR and RESCUE RNA editors by functionalizing Cas13bt with adenosine and cytosine deaminase domains, and demonstrated packaging of the editors within a single adeno-associated virus.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Adenosine / genetics
Adenosine Deaminase / genetics
Animals
CRISPR-Cas Systems* / genetics
Gene Editing
Mammals / genetics
RNA Editing / genetics
RNA* / genetics

Substances

RNA
Adenosine Deaminase
Adenosine

Abstract

Publication types

MeSH terms

Substances

Grants and funding