Using a barcoded AAV capsid library to select for clinically relevant gene therapy vectors

JCI Insight. 2019 Nov 14;4(22):e131610. doi: 10.1172/jci.insight.131610.


While gene transfer using recombinant adeno-associated viral (rAAV) vectors has shown success in some clinical trials, there remain many tissues that are not well transduced. Because of the recent success in reprogramming islet-derived cells into functional β cells in animal models, we constructed 2 highly complex barcoded replication competent capsid shuffled libraries and selected for high-transducing variants on primary human islets. We describe the generation of a chimeric AAV capsid (AAV-KP1) that facilitates transduction of primary human islet cells and human embryonic stem cell-derived β cells with up to 10-fold higher efficiency compared with previously studied best-in-class AAV vectors. Remarkably, this chimeric capsid also enabled transduction of both mouse and human hepatocytes at very high levels in a humanized chimeric mouse model, thus providing a versatile vector that has the potential to be used in both preclinical testing and human clinical trials for liver-based diseases and diabetes.

Keywords: Diabetes; Embryonic stem cells; Gene therapy; Therapeutics.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Capsid / chemistry*
  • Cells, Cultured
  • Dependovirus / genetics*
  • Diabetes Mellitus
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism
  • Gene Library
  • Gene Transfer Techniques
  • Genetic Therapy / methods*
  • Genetic Vectors / genetics*
  • HEK293 Cells
  • Hepatocytes / cytology
  • Hepatocytes / metabolism
  • Humans
  • Islets of Langerhans / cytology
  • Islets of Langerhans / metabolism
  • Mice