Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Robust RNA editing via recruitment of endogenous ADARs using circular guide RNAs

Dhruva Katrekar, James Yen, Yichen Xiang, Anushka Saha, Dario Meluzzi, Yiannis Savva, Prashant Mali
doi: https://doi.org/10.1101/2021.01.12.426286
Dhruva Katrekar
1Department of Bioengineering, University of California San Diego, CA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
James Yen
1Department of Bioengineering, University of California San Diego, CA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Yichen Xiang
1Department of Bioengineering, University of California San Diego, CA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Anushka Saha
1Department of Bioengineering, University of California San Diego, CA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Dario Meluzzi
2Department of Medicine, University of California San Diego, CA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Yiannis Savva
3Shape Therapeutics, WA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Prashant Mali
1Department of Bioengineering, University of California San Diego, CA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: pmali@ucsd.edu
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Preview PDF
Loading

ABSTRACT

Akin to short-hairpin RNAs and antisense oligonucleotides which efficaciously recruit endogenous cellular machinery such as Argonaute and RNase H to enable targeted RNA knockdown, simple long antisense guide RNAs (1) can recruit endogenous adenosine deaminases acting on RNA (ADARs) to enable programmable A-to-I RNA editing, without requiring co-delivery of any exogenous proteins. This approach is highly specific, however the efficiency is typically lower than observed with enzyme overexpression. Conjecturing this was due in part to the short half-life and residence times of guide RNAs, here we engineer highly stable circular ADAR recruiting guide RNAs (cadRNAs), which can be delivered not only by genetically encoding on DNA vectors, but also via transfection of RNA molecules transcribed in vitro. Using these cadRNAs, we observed robust RNA editing across multiple sites and cell lines, in both untranslated and coding regions of RNAs, vastly improved efficiency and durability of RNA editing, and high transcriptome-wide specificity. High transcript-level specificity was achieved by further engineering to reduce bystander editing. Additionally, in vivo delivery of cadRNAs via adeno-associated viruses (AAVs) enabled robust 38% RNA editing of the mPCSK9 transcript in C57BL/6J mice livers, and 12% UAG-to-UGG RNA correction of the amber nonsense mutation in the IDUA-W392X mouse model of mucopolysaccharidosis type I-Hurler (MPS I-H) syndrome. Taken together, cadRNAs enable efficacious programmable RNA editing with application across diverse protein modulation and gene therapeutic settings.

Competing Interest Statement

D.K. and P.M. have filed patents based on this work. P.M. is a scientific co-founder of Shape Therapeutics, Boundless Biosciences, Seven Therapeutics, Navega Therapeutics, and Engine Biosciences. The terms of these arrangements have been reviewed and approved by the University of California, San Diego in accordance with its conflict of interest policies. Y.S. is an employee of Shape Therapeutics.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
Back to top
PreviousNext
Posted January 13, 2021.
Download PDF
Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Robust RNA editing via recruitment of endogenous ADARs using circular guide RNAs
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Robust RNA editing via recruitment of endogenous ADARs using circular guide RNAs
Dhruva Katrekar, James Yen, Yichen Xiang, Anushka Saha, Dario Meluzzi, Yiannis Savva, Prashant Mali
bioRxiv 2021.01.12.426286; doi: https://doi.org/10.1101/2021.01.12.426286
Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Robust RNA editing via recruitment of endogenous ADARs using circular guide RNAs
Dhruva Katrekar, James Yen, Yichen Xiang, Anushka Saha, Dario Meluzzi, Yiannis Savva, Prashant Mali
bioRxiv 2021.01.12.426286; doi: https://doi.org/10.1101/2021.01.12.426286

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Bioengineering
Subject Areas
All Articles
  • Animal Behavior and Cognition (2540)
  • Biochemistry (4990)
  • Bioengineering (3491)
  • Bioinformatics (15256)
  • Biophysics (6921)
  • Cancer Biology (5414)
  • Cell Biology (7762)
  • Clinical Trials (138)
  • Developmental Biology (4545)
  • Ecology (7171)
  • Epidemiology (2059)
  • Evolutionary Biology (10247)
  • Genetics (7524)
  • Genomics (9813)
  • Immunology (4883)
  • Microbiology (13276)
  • Molecular Biology (5159)
  • Neuroscience (29525)
  • Paleontology (203)
  • Pathology (839)
  • Pharmacology and Toxicology (1469)
  • Physiology (2148)
  • Plant Biology (4772)
  • Scientific Communication and Education (1015)
  • Synthetic Biology (1340)
  • Systems Biology (4016)
  • Zoology (770)