A trial of somatic gene targeting in vivo with an adenovirus vector

Asami Ino, Yasuhiro Naito, Hiroyuki Mizuguchi, Naofumi Handa, Takao Hayakawa, Ichizo Kobayashi

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Background: Gene targeting in vivo provides a potentially powerful method for gene analysis and gene therapy. In order to sensitively detect and accurately measure designed sequence changes, we have used a transgenic mouse system, MutaMouse, which has been developed for detection of mutation in vivo. It carries bacteriophage lambda genome with lacZ+ gene, whose change to lacZ-negative allele is detected after in vitro packaging into bacteriophage particles. We have also demonstrated that gene transfer with a replication-defective adenovirus vector can achieve efficient and accurate gene targeting in vitro. Methods: An 8 kb long DNA corresponding to the bacteriophage lambda transgene with one of two lacZ-negative single-base-pair-substitution mutant allele was inserted into a replication-defective adenovirus vector. This recombinant adenovirus was injected to the transgenic mice via tail-vein. Twenty-four hours later, genomic DNA was extracted from the liver tissue and the lambda::lacZ were recovered by in vitro packaging. The lacZ-negative phage was detected as a plaque former on agar with phenyl-beta-D-galactoside. Results: The mutant frequency of the lacZ-negative recombinant adenovirus injected mice was at the same level with the control mouse (∼1/10000). Our further restriction analysis did not detect any designed recombinant. Conclusions: The frequency of gene targeting in the mouse liver by these recombinant adenoviruses was shown to be less than 1/20000 in our assay. However, these results will aid the development of a sensitive, reliable and PCR-independent assay for gene targeting in vivo mediated by virus vectors and other means.

Original languageEnglish
Article number8
JournalGenetic Vaccines and Therapy
Volume3
DOIs
Publication statusPublished - 2005 Oct 12

Fingerprint

Gene Targeting
Adenoviridae
Bacteriophage lambda
Product Packaging
Bacteriophages
Transgenic Mice
Alleles
Lac Operon
Liver
DNA
Transgenes
Base Pairing
Genetic Therapy
Genes
Agar
Tail
Veins
Genome
Viruses
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

A trial of somatic gene targeting in vivo with an adenovirus vector. / Ino, Asami; Naito, Yasuhiro; Mizuguchi, Hiroyuki; Handa, Naofumi; Hayakawa, Takao; Kobayashi, Ichizo.

In: Genetic Vaccines and Therapy, Vol. 3, 8, 12.10.2005.

Research output: Contribution to journalArticle

Ino, Asami ; Naito, Yasuhiro ; Mizuguchi, Hiroyuki ; Handa, Naofumi ; Hayakawa, Takao ; Kobayashi, Ichizo. / A trial of somatic gene targeting in vivo with an adenovirus vector. In: Genetic Vaccines and Therapy. 2005 ; Vol. 3.
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