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ADP/ATP carrier or AAC protein is the most abundant carrier protein in the inner mitochondrial membrane. It transports large quantities of ADP and ATP, equivalent to the average human body weight, every day. Among other transporters, ACC protein is one of the best-studied members of the mitochondrial carrier protein family. The ADP/ATP carrier protein comprises two transmembrane helices connected to a loop and a single alpha-helix on the matrix side. It switches between two conformational...
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Updated: May 21, 2025

Author Spotlight: Improved Method for Production and Purification of Adeno-Associated Viral Vectors
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Protein Carrier Adeno-Associated Virus.

Mareike Daniela Hoffmann1, Ryan James Sorensen2, Ajay Extross3

  • 1Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota 55455, United States.

ACS Nano
|March 21, 2025
PubMed
Summary
This summary is machine-generated.

Adeno-associated virus (AAV) can now deliver proteins, not just DNA. Researchers engineered AAV to package and deliver various proteins, expanding its gene therapy applications.

Keywords:
adeno-associated viruscapsid engineeringnanoparticlesprotein deliverysynthetic virology

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Production, Purification, and Quality Control for Adeno-associated Virus-based Vectors
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Production, Purification, and Quality Control for Adeno-associated Virus-based Vectors

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Gene Therapy

Background:

  • Adeno-associated virus (AAV) is a key vector for gene therapy, primarily delivering DNA.
  • The capacity of AAV to deliver protein payloads remains largely unexplored.
  • Expanding AAV's utility requires novel delivery mechanisms beyond nucleic acids.

Purpose of the Study:

  • To engineer a protein-carrying AAV (pcAAV) capable of packaging and delivering protein payloads.
  • To investigate the efficiency and factors influencing protein packaging within AAV capsids.
  • To demonstrate the therapeutic potential of AAV-mediated protein delivery.

Main Methods:

  • Engineered AAV capsids by inserting binding domains on the interior surface.
  • Mediated protein packaging via interactions between binding domains and protein tags during capsid assembly.
  • Validated packaging of diverse proteins including fluorescent proteins, Cas9, Cre recombinase, and APEX2.
  • Assessed packaging efficiency based on insertion site, viral protein isoform, and target protein localization.

Main Results:

  • Successfully demonstrated packaging of multiple proteins within engineered AAV capsids.
  • Showcased that packaging efficiency is influenced by specific engineering strategies and protein characteristics.
  • Confirmed successful cellular entry and protein payload delivery by pcAAV.
  • Verified that delivered enzymes retained their biological activity post-packaging.
  • Established that this protein packaging capability is applicable across multiple AAV serotypes.

Conclusions:

  • Adeno-associated virus (AAV) has been successfully engineered as a versatile protein delivery vehicle (pcAAV).
  • This advancement significantly broadens the scope of AAV applications in biomedical research and therapy.
  • pcAAV technology opens new avenues for protein-based therapeutics and diagnostics.