Molecular Design and Production of AAV Viral Vectors for Gene Therapy

Raela B Ridley; Erin M Walsh; Cristhian J Ildefonso

doi:10.1007/978-1-0716-1012-1_5

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.

Authors

Raela B Ridley¹, Erin M Walsh¹, Cristhian J Ildefonso²

Affiliations

¹ Department of Ophthalmology, University of Florida College of Medicine, Gainesville, FL, USA.
² Department of Ophthalmology, University of Florida College of Medicine, Gainesville, FL, USA. ildefons@ufl.edu.

PMID: 33108658
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