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In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
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Reprogramming site-specific retrotransposon activity to new DNA sites
Christopher W Fell1,2,3,4, Lukas Villiger4, Justin Lim4
1Department of Medicine, Division of Engineering in Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.
Nature
|April 9, 2025
Summary
Researchers engineered a new system called STITCHR for precise, scarless genome editing. This retroelement-based tool allows efficient insertion of genetic material at targeted locations in both dividing and non-dividing cells.
Area of Science:
- Genomics and Molecular Biology
- Retrotransposon Biology
- Gene Editing Technologies
Background:
- Non-long terminal repeat (non-LTR) retrotransposons are key drivers of eukaryotic genome evolution.
- These mobile genetic elements often integrate into specific repetitive genomic regions.
- The precise targeting mechanisms and limitations of retrotransposons remain incompletely understood.
Purpose of the Study:
- To discover and characterize new site-specific retrotransposon families.
- To investigate the insertion preferences and retargeting potential of retrotransposons.
- To engineer a novel platform for precise and scarless genomic integration.
Main Methods:
- Utilized a computational pipeline to identify novel retrotransposon families.
- Performed biochemical and cellular profiling of identified retrotransposon members.
- Engineered a retrotransposon-CRISPR fusion system (STITCHR) for targeted insertion.
Main Results:
- Discovered new site-specific retrotransposon families with novel insertion preferences.
- Successfully retargeted an R2 retrotransposon (R2Tg) for scarless insertion of payloads.
- Developed STITCHR, enabling efficient, scarless installation of edits up to 12.7 kb, gene replacement, and RNA template use.
Conclusions:
- STITCHR represents a versatile platform for scarless, programmable genome engineering.
- The system demonstrates potential for applications in both research and therapeutic settings.
- This approach leverages the natural prevalence of non-LTR retrotransposons for advanced gene editing.
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