FVB;B6-Rnf41^{TgTn(sb-rtTA,-Tyr)2435COve}/Mmjax

Stock number: 017421
Research areas: Reproductive Biology Research

Description

These OVE#2435C mice harbor a mutation created by random insertion of the SB-sa-IRES-rtTA-pA-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2223). The donating investigator reports the phenotype of homozygous mice as: males are sterile.

Detailed description

These OVE#2435C mice harbor a mutation created by random insertion of the SB-sa-IRES-rtTA-pA-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2223). Using inverse PCR analysis, the transgene integration site was identified in intron 2 of the ring finger protein 41 gene (Rnf41) on chromosome 10 (specifically at the 5'-127,864,981(+) bp position with the rtTA in sense orientation relative to the endogenous gene). The donating investigator reports the phenotype of homozygous mice as: males are sterile.

Development

A lentiviral transgenic approach was used to generate these mice. The SB-sa-IRES-rtTA-pA-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2223) was designed with a Sleeping Beauty (SB) transposon (containing a slice-acceptor::IRES::rtTA::polyA gene trap), a mouse tyrosinase minigene (Tyro), and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) in the FUGW self-inactivating HIV-based lentiviral vector backbone. The SB transposon and Tyro minigene replaced the ubiquitin-c promoter and EGFP sequences originally found in the FUGW lentiviral vector. The SB transposon used in this transgene has an inverted repeat/direct repeat sequence (IR/DR; the SB transposon recognition site), an adenovirus splice acceptor, an internal ribosome entry site (IRES; from human X-chromosome-linked inhibitor of apoptosis (XIAP)), a reverse tetracycline controlled transactivator sequence (rtTA; with stop codon), a human growth hormone polyA sequence, and a second IR/DR sequence. The IR/DR sequences are outward-facing (pointed away from the sa-IRES-rtTA-pA). The Tyro minigene is composed of the mouse Tyr enhancer region (620 bp), promoter region (500 bp), and 1.7 kbp cDNA sequence (including the stop codon); all in sense orientation relative to the FUGW backbone. There is no polyA site between the Tyro minigene and WPRE sequence. The WPRE sequence functions to enhance the mRNA transcript stability. This packaged lentiposon construct was injected subzonally (under the zona pellucida) into 1-8 cell stage C57BL/6 mouse embryos. Founder mice (F0) with random, multiple lentiviral integration sites were identified by PCR. Founder mice from line OVE#2435C were bred with FVB/N mice, and transgenic offspring (F1) were bred to FVB/N mice. The resulting transgenic offspring (F2) with identical coat colors (light grey) were subsequently inbred. Using inverse PCR analysis, the transgene integration site was identified in intron 2 of the ring finger protein 41 gene (Rnf41) on chromosome 10 (specifically at the 5'-127,864,981(+) bp position with the rtTA in sense orientation relative to the endogenous gene). Transgenic males were sent to The Jackson Laboratory Repository (MMRRC-JAX). Upon arrival, these mice were bred to FVB/NJ inbred mice for at least one generation to establish the live colony.

Allele

Symbol: Rnf41^{TgTn(sb-rtTA,Tyr)2435COve}
Name: transgenic transposon lentiviral insertion 2435C, Paul A Overbeek
MGI accession ID: MGI:5287504
Generation method: Transgenic
Attributes: Transposon concatemer
Synonyms: OVE#2435; OVE2435
Marker symbol: Rnf41
Marker name: ring finger protein 41
Marker synonyms: 2210404G21Rik; 4930511A05Rik; 4933415P08Rik; D10Ertd722e; FLRF; Nrdp1; SBBI03
Chromosome: 10
Strain of origin: C57BL/6
Expressed genes: Tyr,tyrosinase,mouse, laboratory
Molecular note: A lentiviral transgenic approach was used to generate these mice. The SB-sa-IRES-rtTA-pA-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2223) was designed with a Sleeping Beauty (SB) transposon (containing a slice-acceptor::IRES::rtTA::polyA gene trap), a mouse tyrosinase minigene (Tyro), and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) in the FUGW self-inactivating HIV-based lentiviral vector backbone. The SB transposon and Tyro minigene replaced the ubiquitin-c promoter and EGFP sequences originally found in the FUGW lentiviral vector. The 1200 bp SB transposon used in this transgene has an inverted repeat/direct repeat sequence (IR/DR; the SB transposon recognition site), an adenovirus splice acceptor, a stop sequence (3xSTOP), an internal ribosome entry site (IRES; from human X-chromosome-linked inhibitor of apoptosis (XIAP)), a sequence encoding a-n optimized form of reverse tetracycline controlled transactivator (rtTA2S; with stop codon), a human growth hormone polyA sequence, and a second IR/DR sequence. The IR/DR sequences are outward-facing (pointed away from the sa-IRES-rtTA-pA). The Tyro minigene is composed of the mouse Tyr enhancer region (623 bp), promoter region (657 bp), and 1566 bp cDNA sequence (including the stop codon); all in sense orientation relative to the FUGW backbone. There is no polyA site between the Tyro minigene and WPRE sequence. The WPRE sequence functions to enhance the mRNA transcript stability. In line OVE2435C, a single copy of the lentiposon inserted into intron 2 [NCBI37/mm9; 5'-127,864,981(+)] in the sense orientation.