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The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Updated: Jul 15, 2025

Engineering and Evolution of Synthetic Adeno-Associated Virus AAV Gene Therapy Vectors via DNA Family Shuffling
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Double-Strand Break Repair Pathways Differentially Affect Processing and Transduction by Dual AAV Vectors.

Anna C Maurer1,2, Brian Benyamini1, Vinson B Fan1

  • 1Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.

Biorxiv : the Preprint Server for Biology
|October 4, 2023
PubMed
Summary
This summary is machine-generated.

Inhibiting DNA repair pathways like Homologous Recombination (HR) enhances gene delivery using dual recombinant adeno-associated viral vectors (rAAV). This improves the expression of large transgenes, overcoming rAAV size limitations.

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Area of Science:

  • Molecular Biology
  • Gene Therapy
  • Virology

Background:

  • Recombinant adeno-associated viral vectors (rAAV) are widely used for gene delivery.
  • Current rAAV vectors have limited DNA carrying capacity, restricting therapeutic applications.
  • Dual trans-splicing vectors expand payload by concatenating rAAV genomes, but efficiency is variable.

Conclusions:

  • Host DNA damage repair pathways, specifically HR, play a critical role in rAAV transduction efficiency.
  • Targeting HR factors offers a novel strategy to improve gene payload capacity of rAAV vectors.
  • Pharmacological inhibition of HR presents a promising approach for enhancing gene therapy delivery.