Abundant off-target edits from site-directed RNA editing can be reduced by nuclear localization of the editing enzyme

Isabel C. Vallecillo-Viejo, Noa Liscovitch-Brauer, Maria Fernanda Montiel-Gonzalez, Eli Eisenberg, Joshua J.C. Rosenthal

Research output: Contribution to journalArticlepeer-review

65 Scopus citations


Site-directed RNA editing (SDRE) is a general strategy for making targeted base changes in RNA molecules. Although the approach is relatively new, several groups, including our own, have been working on its development. The basic strategy has been to couple the catalytic domain of an adenosine (A) to inosine (I) RNA editing enzyme to a guide RNA that is used for targeting. Although highly efficient on-target editing has been reported, off-target events have not been rigorously quantified. In this report we target premature termination codons (PTCs) in messages encoding both a fluorescent reporter protein and the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein transiently transfected into human epithelial cells. We demonstrate that while on-target editing is efficient, off-target editing is extensive, both within the targeted message and across the entire transcriptome of the transfected cells. By redirecting the editing enzymes from the cytoplasm to the nucleus, off-target editing is reduced without compromising the on-target editing efficiency. The addition of the E488Q mutation to the editing enzymes, a common strategy for increasing on-target editing efficiency, causes a tremendous increase in off-target editing. These results underscore the need to reduce promiscuity in current approaches to SDRE.

Original languageEnglish
Pages (from-to)104-114
Number of pages11
JournalRNA Biology
Issue number1
StatePublished - 2 Jan 2018
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2018 Taylor & Francis Group, LLC.


  • ADAR
  • RNA editing
  • off-target RNA editing
  • site-directed RNA editing


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