Viral serotypes have evolved diverse tissue tropisms through natural selection and an iterative process of genetic recombination and mutagenesis. Within this framework, the recent identification of Adeno-Associated Virus (AAV) serotypes with broad tissue tropisms has provided the gene therapy community with a versatile toolkit of AAV vectors. Our goal is to establish a thorough understanding of the structure-function correlates of the diverse tissue tropisms of AAV serotypes. To achieve such, we have devised a comprehensive, two-pronged approach to unravel structural attributes of AAV1-9, while simultaneously exploiting these serotypes as """"""""blueprints"""""""" for novel AAV vector design. The approach exploits the ability of DNA shuffling to rapidly evolve novel phenotypes derived from parental serotypes followed by rational manipulation of novel AAV mutants to establish structural attributes at the amino acid level. The first strategy involves generation of a combinatorial AAV library through DNA shuffling of AAV serotype capsid sequences followed by directed evolution of cell type/receptor-specific mutants. The objective of this approach is to eliminate bias in the identification of so-called """"""""hot spots"""""""" on the AAV capsid that impart a specific phenotype. The second approach is concerned with rational manipulation of such specific regions on AAV serotype capsids using site-directed mutagenesis. The goal of the latter strategy is to establish structure-function correlates of the AAV capsid at the amino acid level. The two complementary strategies are expected to generate critical structural information that will lay the groundwork for custom design of tissue-targeted AAV vectors. Research design involves the (1) synthesis of an AAV library through aforementioned strategies, directed evolution of cell type/receptor-specific AAV mutants, (2) characterization of such novel variants using molecular modeling, cryo-EM, a battery of biochemical assays in vitro, and (3) their translation into vectors for independent gene delivery applications in vivo.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Project (R01)
Project #
Application #
Study Section
Gene and Drug Delivery Systems Study Section (GDD)
Program Officer
Park, Eun-Chung
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of North Carolina Chapel Hill
Schools of Medicine
Chapel Hill
United States
Zip Code
Song, Liujiang; Llanga, Telmo; Conatser, Laura M et al. (2018) Serotype survey of AAV gene delivery via subconjunctival injection in mice. Gene Ther 25:402-414
Chai, Zheng; Zhang, Xintao; Rigsbee, Kelly Michelle et al. (2018) Cryoprecipitate augments the global transduction of the adeno-associated virus serotype 9 after a systemic administration. J Control Release 286:415-424
Shao, Wenwei; Chen, Xiaojing; Samulski, Richard J et al. (2018) Inhibition of antigen presentation during AAV gene therapy using virus peptides. Hum Mol Genet 27:601-613
Shao, Wenwei; Earley, Lauriel F; Chai, Zheng et al. (2018) Double-stranded RNA innate immune response activation from long-term adeno-associated virus vector transduction. JCI Insight 3:
Chai, Zheng; Samulski, R Jude; Li, Chengwen (2018) Nab Escaping AAV Mutants Isolated from Mouse Muscles. Bio Protoc 8:
Brown, Nolan; Song, Liujiang; Kollu, Nageswara R et al. (2017) Adeno-Associated Virus Vectors and Stem Cells: Friends or Foes? Hum Gene Ther 28:450-463
Llanga, Telmo; Nagy, Nadia; Conatser, Laura et al. (2017) Structure-Based Designed Nano-Dysferlin Significantly Improves Dysferlinopathy in BLA/J Mice. Mol Ther 25:2150-2162
Hirsch, Matthew L; Conatser, Laura M; Smith, Sara M et al. (2017) AAV vector-meditated expression of HLA-G reduces injury-induced corneal vascularization, immune cell infiltration, and fibrosis. Sci Rep 7:17840
Nagy, Nadia; Nonneman, Randal J; Llanga, Telmo et al. (2017) Hip region muscular dystrophy and emergence of motor deficits in dysferlin-deficient Bla/J mice. Physiol Rep 5:
Liang, Katharine J; Woodard, Kenton T; Weaver, Mark A et al. (2017) AAV-Nrf2 Promotes Protection and Recovery in Animal Models of Oxidative Stress. Mol Ther 25:765-779

Showing the most recent 10 out of 65 publications