Molecular Design and Production of AAV Viral Vectors for Gene Therapy

Methods Mol Biol. 2021:2225:77-92. doi: 10.1007/978-1-0716-1012-1_5.

Abstract

Adeno-associated virus (AAV) is a helper-dependent single-stranded DNA parvovirus. Over the years, AAV has become the vector of choice in the gene therapy field due to its safety profile and low immunogenicity. With a carrying capacity of 4.2 kbp, these vectors have demonstrated their clinical value, especially in the field of ophthalmology. Herein we describe methods for the molecular design and packaging of AAV viral vectors. These methods apply to the design of single-stranded or self-complementary AAV vectors.

Keywords: AAV; Cloning; Gene therapy; Ligation; PCR; Transformation; Transgene.

Publication types

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

MeSH terms

  • Cloning, Molecular / methods*
  • DNA Primers / chemistry
  • DNA Primers / metabolism
  • DNA, Single-Stranded / genetics
  • DNA, Single-Stranded / metabolism
  • Dependovirus / genetics*
  • Dependovirus / metabolism
  • Escherichia coli / virology
  • Genetic Engineering / methods*
  • Genetic Therapy / methods
  • Humans
  • Plasmids / chemistry
  • Plasmids / metabolism
  • Transduction, Genetic
  • Transgenes*
  • Viral Genome Packaging / genetics*

Substances

  • DNA Primers
  • DNA, Single-Stranded