An engineered U7 small nuclear RNA scaffold greatly increases ADAR-mediated programmable RNA base editing
Abstract Custom RNA base editing exploiting the human Adenosine Deaminase Acting on RNA (ADAR) enzyme may enable therapeutic gene editing without DNA damage or use of foreign proteins. ADAR’s adenosine-to-inosine (effectively A-to-G) deamination activity can be targeted to transcripts using an antis...
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| Format: | Article |
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Nature Portfolio
2025-05-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-60155-z |
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| author | Susan M. Byrne Stephen M. Burleigh Robert Fragoza Yue Jiang Yiannis Savva Ricky Pabon Evan Kania Joseph Rainaldi Andrew Portell Prashant Mali Adrian W. Briggs |
| author_facet | Susan M. Byrne Stephen M. Burleigh Robert Fragoza Yue Jiang Yiannis Savva Ricky Pabon Evan Kania Joseph Rainaldi Andrew Portell Prashant Mali Adrian W. Briggs |
| author_sort | Susan M. Byrne |
| collection | DOAJ |
| description | Abstract Custom RNA base editing exploiting the human Adenosine Deaminase Acting on RNA (ADAR) enzyme may enable therapeutic gene editing without DNA damage or use of foreign proteins. ADAR’s adenosine-to-inosine (effectively A-to-G) deamination activity can be targeted to transcripts using an antisense guide RNA (gRNA), but efficacy is challenged by limits of in vivo delivery. Embedding gRNAs into a U7 small nuclear RNA (snRNA) framework greatly enhances RNA editing with endogenous ADAR, and a 750-plex single-cell mutagenesis screen further improved the framework. An optimized scaffold with a stronger synthetic U7 promoter enables 76% RNA editing in vitro from a single DNA construct per cell, and 75% editing in a Hurler syndrome mouse brain after one systemic AAV injection, surpassing circular gRNA approaches. The technology also improves published DMD exon-skipping designs 25-fold in differentiated myoblasts. Our engineered U7 framework represents a universal scaffold for ADAR-based RNA editing and other antisense RNA therapies. |
| format | Article |
| id | doaj-art-24d57bfe48074d40ab3ea830e45a5b9f |
| institution | DOAJ |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-24d57bfe48074d40ab3ea830e45a5b9f2025-08-20T03:22:03ZengNature PortfolioNature Communications2041-17232025-05-0116111410.1038/s41467-025-60155-zAn engineered U7 small nuclear RNA scaffold greatly increases ADAR-mediated programmable RNA base editingSusan M. Byrne0Stephen M. Burleigh1Robert Fragoza2Yue Jiang3Yiannis Savva4Ricky Pabon5Evan Kania6Joseph Rainaldi7Andrew Portell8Prashant Mali9Adrian W. Briggs10Shape TherapeuticsShape TherapeuticsShape TherapeuticsShape TherapeuticsShape TherapeuticsShape TherapeuticsShape TherapeuticsUniversity of California San DiegoUniversity of California San DiegoUniversity of California San DiegoShape TherapeuticsAbstract Custom RNA base editing exploiting the human Adenosine Deaminase Acting on RNA (ADAR) enzyme may enable therapeutic gene editing without DNA damage or use of foreign proteins. ADAR’s adenosine-to-inosine (effectively A-to-G) deamination activity can be targeted to transcripts using an antisense guide RNA (gRNA), but efficacy is challenged by limits of in vivo delivery. Embedding gRNAs into a U7 small nuclear RNA (snRNA) framework greatly enhances RNA editing with endogenous ADAR, and a 750-plex single-cell mutagenesis screen further improved the framework. An optimized scaffold with a stronger synthetic U7 promoter enables 76% RNA editing in vitro from a single DNA construct per cell, and 75% editing in a Hurler syndrome mouse brain after one systemic AAV injection, surpassing circular gRNA approaches. The technology also improves published DMD exon-skipping designs 25-fold in differentiated myoblasts. Our engineered U7 framework represents a universal scaffold for ADAR-based RNA editing and other antisense RNA therapies.https://doi.org/10.1038/s41467-025-60155-z |
| spellingShingle | Susan M. Byrne Stephen M. Burleigh Robert Fragoza Yue Jiang Yiannis Savva Ricky Pabon Evan Kania Joseph Rainaldi Andrew Portell Prashant Mali Adrian W. Briggs An engineered U7 small nuclear RNA scaffold greatly increases ADAR-mediated programmable RNA base editing Nature Communications |
| title | An engineered U7 small nuclear RNA scaffold greatly increases ADAR-mediated programmable RNA base editing |
| title_full | An engineered U7 small nuclear RNA scaffold greatly increases ADAR-mediated programmable RNA base editing |
| title_fullStr | An engineered U7 small nuclear RNA scaffold greatly increases ADAR-mediated programmable RNA base editing |
| title_full_unstemmed | An engineered U7 small nuclear RNA scaffold greatly increases ADAR-mediated programmable RNA base editing |
| title_short | An engineered U7 small nuclear RNA scaffold greatly increases ADAR-mediated programmable RNA base editing |
| title_sort | engineered u7 small nuclear rna scaffold greatly increases adar mediated programmable rna base editing |
| url | https://doi.org/10.1038/s41467-025-60155-z |
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