How do LINE family retrotransposons jump around?
Can we reprogram retrotransposons?
What does this have to do with Rum and Raisin?
Answers here and below!
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R2 is an exquisite LINE-like retrotransposon that inserts in ribosomal DNA repeats. Thomas Eickbush et al have done so much awesome work, showing that the R2 protein is INSANE: by itself, it finds a DNA target, cuts it, and reverse transcribes its own RNA right there and then.
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We had a bunch of questions about R2, so solved its structure #CryoEM
Q: How does R2 find its target?
A: We found two sequence motifs, we call them the RUM and RASIN 🥃+🍇
RUM is bound by accessory DNA binding domains on R2.
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When R2 binds the RUM, it unzips the RASIN, then cuts one strand of the RASIN.
The cut strand is used as a primer for reverse transcription of the R2 RNA - so this is called TPRT (target-primed reverse transcription)
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Now it gets really fun: based on the structure, we can devise the ✨R2-tag✨ - 43 nt that you can attach to any RNA, and R2 will integrate it into the ribosomal DNA.
(sorry, I ran out of movies)
Apr 6, 2023 · 6:25 PM UTC
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ALSO R2 doesn't have to use the ribosomal DNA target, it can be retargeted: fused to Cas9, it can instead reverse transcribe at the Cas9 nick site. Maybe this could one day be really useful for gene therapy.
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(Just kidding, one more movie)
Thank you my lovely co-authors @FrangiehChris @zhangf and Rhi Macrae
Also @bradyajohnston's #MolecularNodes Blender plugin is great. Everyone should use it. #b3d
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the inevitable forgotten link to the actual paper!
science.org/doi/10.1126/scie…
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