Self-complementary recombinant adeno-associated viral vectors: packaging capacity and the role of rep proteins in vector purity.

Self-complementary recombinant adeno-associated viral vectors: packaging capacity and the role of rep proteins in vector purity. Research Abstract Details 

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Self-complementary recombinant adeno-associated viral vectors: packaging capacity and the role of rep proteins in vector purity. Abstract Text:

Jianqing Wu,Weihong Zhao,Li Zhong,Zongchao Han,Baozheng Li,Wenqin Ma,Kirsten A Weigel-Kelley,Kenneth H Warrington,Arun Srivastava,

Self-complementary adeno-associated viral (scAAV) vectors bypass the requirement for viral second-strand DNA synthesis, but the packaging capacity of these vectors ( approximately 2.4 kb) is significantly smaller than that of conventional AAV vectors ( approximately 4.8 kb). We constructed human recombinant green fluorescent protein (hrGFP) expression cassettes ranging from 2.3 to 4.1 kb. Each vector was biologically active, but the transduction efficiency of vectors containing <3.3-kb genomes was significantly higher than those containing 3.5-kb genomes or larger. However, scAAV vectors containing up to approximately 3.3-kb genomes also contained single-stranded genomes, and 3.5-kb and larger genomes were packaged only as single-stranded DNA. These data suggest that the maximum packaging capacity of scAAV vectors is approximately 3.3 kb. The production of single-stranded genomes was not due to repair of the terminal resolution site (trs) in the inverted terminal repeats in the AAV genome, but rather was partly due to the use of AAV helper plasmid, known to lead to higher levels of expression of Rep proteins. The use of a helper plasmid known to lead to reduced levels of Rep proteins led to the generation of scAAV vectors that contained approximately 90% of the viral genomes in double-stranded forms. These studies demonstrate the feasibility of achieving encapsidation of larger genomes into scAAV vectors than was suggested originally, but underscore the need to exercise caution in using the appropriate helper plasmid to generate scAAV stocks capable of high-efficiency transduction that are relatively free of single-stranded DNA-containing vectors.

Self-complementary recombinant adeno-associated viral vectors: packaging capacity and the role of rep proteins in vector purity. Publishing Authors By Initials

J Wu,W Zhao,L Zhong,Z Han,B Li,W Ma,KA Weigel-Kelley,KH Warrington,A Srivastava,

For similar biological phenomena, cell phenomena, and immunity: biological phenomena: microbiologic phenomena: viral physiology: virus replication research abstracts see: biological phenomena, cell phenomena, and immunity: biological phenomena: microbiologic phenomena: viral physiology: virus replication research

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Self-complementary recombinant adeno-associated viral vectors: packaging capacity and the role of rep proteins in vector purity. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Human gene therapy

VOLUME: 18

Page Numbers: 171-82

Journal Abbreviation: Hum. Gene Ther.

ISSN: 1043-0342

DAY: 3

MONTH: Feb

YEAR: 2007

Self-complementary recombinant adeno-associated viral vectors: packaging capacity and the role of rep proteins in vector purity. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 9008950

Self-complementary recombinant adeno-associated viral vectors: packaging capacity and the role of rep proteins in vector purity. Keywords Mesh Terms:

KEYWORDS: Virus Replication

MESH TERMS: physiology

Chemical & Substance for Abstract: Self-complementary recombinant adeno-associated viral vectors: packaging capacity and the role of rep proteins in vector purity. Information

Substance Name: rep proteins, Adeno-associated virus 2

Registry Number: 137750-19-7

Grant and Affiliation Information for Self-complementary recombinant adeno-associated viral vectors: packaging capacity and the role of rep proteins in vector purity.

AFFILIATION: Division of Cellular & Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL 32610, USA.

Country: United States

AGENCY: United States NHLBI

GRANT: R01 HL-76901

ACRONYM: HL

MEDLINETA: Hum Gene Ther

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