C3;B6-Tg(AAVS1)A1Xob/J

Stock number: 009601
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools, Virology Research

Description

Hemizygous mice are viable and fertile with no reported phenotypic abnormalities from the insertion of five full-length human AAVS1 sites (adeno-associated virus integration site 1) at a single chromosomal locus. The AAVS1 loci within the transgene allow site-specific integration of hybrid vectors containing AAV inverted terminal repeat-flanked genes of interest.

Detailed description

Hemizygous mice are viable and fertile with no reported phenotypic abnormalities from the insertion of multiple full-length human AAVS1 sites (adeno-associated virus integration site 1) at a single chromosomal locus. The multiple (approximately five) AAVS1 loci within the transgene allow site-specific integration of hybrid vectors containing AAV inverted terminal repeat-flanked genes of interest.

Adeno-associated virus (AAV) is a nonpathogenic parvovirus with a single-stranded DNA genome (4.7 kb) containing two genes (rep and cap) flanked by 145-base inverted terminal repeats (ITRs). Efficient replication of the AAV genome occurs in the presence of a helper virus, either adenovirus or herpes simplex virus. In the absence of a helper virus, ITRs and Rep proteins are sufficient to mediate the integration of AAV sequences into a specific AAVS1 site in the host cell genome and thereby establish a latent infection. In addition, low levels of helper-independent AAV replication can also occur in cells that are chemically synchronized to the S phase or exposed to toxic conditions, including UV irradiation, heat, or carcinogens. Because of this, the AAVS1 loci within the transgene are a platform for site-specific integration of AAV ITR-flanked hybrid vectors that contain genes of interest.

Development

An 8.2 kb human genomic fragment containing the entire adeno-associated virus integration site 1 (AAVS1) locus was isolated after EcoRI digestion of plasmid pRIA-N3. This transgene was microinjected into fertilized oocytes of B6C3F1 mice. Male founders were bred with wildtype C3H female mice (C3H substrain not known). Transgenic founder line A1 was established and found to have approximately 5 copies of the transgene integrated at a single locus. The AAVS1 transgenic mice were maintained by breeding together for many generations prior to arrival at The Jackson Laboratory. Upon arrival, mice were bred with C3HeB/FeJ inbred mice (Stock No. 000658) for at least one generation to establish the colony.

Allele

Symbol: Tg(AAVS1)A1Xob
Name: transgene insertion A1, Xandra Breakefield
MGI accession ID: MGI:3852016
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Tg(AAVS1)A1Xob
Marker name: transgene insertion A1, Xandra Breakefield
Chromosome: UN
Strain of origin: (C57BL/6 x C3H)F1
Expressed genes: AAVS1,adeno-associated virus integration site 1,human
Molecular note: An 8.2 kb human genomic fragment containing the entire adeno-associated virus integration site 1 (AAVS1) locus was isolated after EcoRI digestion of plasmid pRIA-N3. Transgenic founder line A1 was established and found to have approximately 5 copies of the transgene integrated at a single locus.