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Protein Carrier AAV.

Mareike D Hoffmann1, Ryan J Sorensen2, Ajay Extross3

  • 1Department of Genetics, Cell Biology & Development, University of Minnesota, Minneapolis, MN, 55455, USA.

Biorxiv : the Preprint Server for Biology
|August 26, 2024
PubMed
Summary
This summary is machine-generated.

Researchers engineered Adeno-Associated Virus (AAV) capsids to directly package proteins, creating Protein Carrier AAV (pcAAV). This novel platform successfully delivers active protein payloads like GFP and Cas9 into cells, expanding AAV applications.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Gene Therapy

Background:

  • Adeno-Associated Virus (AAV) is a common vector for DNA delivery.
  • The potential of AAV capsids for direct protein payload delivery remains largely unexplored.

Purpose of the Study:

  • To engineer AAV capsids capable of directly packaging functional proteins.
  • To establish Protein Carrier AAV (pcAAV) as a novel protein delivery platform.

Main Methods:

  • Engineered AAV capsids by inserting nanobodies into the capsid interior.
  • Mediated packaging of various proteins including GFP, Cas9, Cre recombinase, and APEX2.
  • Optimized protein packaging by manipulating nanobody insertion site, capsid isoform, and subcellular localization.

Main Results:

  • Demonstrated successful packaging of diverse protein payloads within engineered AAV capsids.
  • Showcased that packaged proteins retain enzymatic activity post-packaging.
  • Confirmed pcAAV's ability to bind, enter cells, and deliver functional protein cargo.
  • Identified key factors influencing protein packaging efficiency for rational engineering.

Conclusions:

  • Protein Carrier AAV (pcAAV) represents a breakthrough in protein delivery technology.
  • pcAAV expands the utility of AAV vectors beyond DNA delivery for research and therapeutics.
  • This platform holds promise for advancing gene therapy and molecular research tools.