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In vivo expansion of gene-targeted hepatocytes through transient inhibition of an essential gene.

Marco De GiorgiSo Hyun ParkAdam CastorenoMingming CaoAyrea HurleyLavanya SaxenaMarcel A ChuecosChristopher J WalkeyAlexandria M DoerflerMia N FurgursonM Cecilia LjungbergKalyani R PatelSarah HydeTyler ChickeringStephanie LefebvreKelly WassarmanPatrick MillerJune QinMark K SchlegelIvan ZlatevRich Gang LiJong KimJames F MartinKarl-Dimiter BissigVasant JadhavGang BaoWilliam R Lagor
Published in: bioRxiv : the preprint server for biology (2023)
Homology Directed Repair (HDR)-based genome editing is an approach that could permanently correct a broad range of genetic diseases. However, its utility is limited by inefficient and imprecise DNA repair mechanisms in terminally differentiated tissues. Here, we tested "Repair Drive", a novel method for improving targeted gene insertion in the liver by selectively expanding correctly repaired hepatocytes in vivo . Our system consists of transient conditioning of the liver by knocking down an essential gene, and delivery of an untargetable version of the essential gene in cis with a therapeutic transgene. We show that Repair Drive dramatically increases the percentage of correctly targeted hepatocytes, up to 25%. This resulted in a five-fold increased expression of a therapeutic transgene. Repair Drive was well-tolerated and did not induce toxicity or tumorigenesis in long term follow up. This approach will broaden the range of liver diseases that can be treated with somatic genome editing.
Keyphrases
  • genome editing
  • crispr cas
  • copy number
  • genome wide
  • dna repair
  • genome wide identification
  • dna damage
  • poor prognosis
  • cancer therapy
  • gene expression
  • liver injury
  • long non coding rna
  • drug delivery
  • dna damage response