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| Strain Number: | 01XGB | ||
| Common Strain Name: | Rosa LSL SBase/TG12740 | ||
| Strain Nomenclature: | STOCK Gt(ROSA)26Sortm2(sb11)Njen TgTn(sb-T2/Onc3)12740Njen | ||
| Release Category (Required for MTA form): C1 | |||
| Animal Health Report in PDF Format | |||
| Strain Description: | This strain is double homozygous for both the TgTn(sb-T2/Onc3)12740Njen (TG12740) Sleeping Beauty mutagenic transgene and a new cre-inducible SBase knock-in allele at ROSA26. This double homozygous strain has a genotype designation of lsl-SBase/lsl-SBase; SB/SB, where lsl-SBase refers to the cre-inducible SBase allele at ROSA26 and SB refers to the TG12740 transgene. Homozygosity for the cre-inducible SBase allele at ROSA26 was determined by PCR genotyping and homozygosity for the TG12740 transgene was determined by a combination of PCR and FISH genotyping; both alleles were confirmed homozygous by test crossing to wild-type mice. A major advantage of the bi-partite Sleeping Beauty mutagenesis system is the ability to selectively control transposase (SBase) expression both spatially and temporally. To create the cre-inducible SBase knock-in allele ROSA26, the same approach was taken as before (see NCI Mouse Repository Strain 01XBG), except that a floxed-STOP (loxP-STOP-loxP) cassette was inserted upstream of the SBase (SB11) gene, thereby preventing SBase expression. Tissue-specific SB expression may be controlled easily by crossing with various cre-expressing mice. Importantly, after deletion of the floxed-STOP cassestte by cre-loxP recombination, SB functions in the same manner as the constitutive SBase allele (NCI Mouse Repository Strain Number 01XBG). The Sleeping Beauty mutagenic transposon [Tn(sb-T2/Onc3); T2/Onc3] was engineered to contain an engrailed-2 splice acceptor flanked by SB transposase binding sites that were optimized to increase transposition frequency. The transposon also contains splice acceptors and a bi-directional polyadenylation signal that can terminate transcription when integrated in either orientation into a gene. A CMV enhancer/chicken ß-actin promoter that drives oncogene expression and a splice donor were included to activate gene expression when integrated upstream of a gene. Two strains of transgenic T2/Onc3 mice were generated: strain TG12740 [B6;C3- TgTn(sb-T2/Onc3)12740Njen] contains a transposon concatemer on chromosome 9. This double homozygous strain is part of the next generation Sleeping Beauty mutagenesis system developed in Nancy Jenkins' laboratory. | ||
| Mutation Information | |||
| Mutation Type: | Targeted | ||
| Gene Name: | ROSA26, sleeping beauty transposon (mouse) | ||
| Gene Symbol: | ROSA26, sb | ||
| Transgene Name: | |||
| Transgene Symbol: | |||
| Promoter Name: | , CAG (chicken) | ||
| Promoter Symbol: | , CAG | ||
| Current Genetic Background: | STOCK | ||
| Approx. Generation: | 4 | ||
| Organ Site : | Various | ||
| Husbandry: | Homozygote breeder pairs are provided. Average litter size is 6. Strain is maintained as homozygous and cannot be readily genotyped by PCR. It can, however, be genotyped by FISH analysis. | ||
| Genotyping Information: | There are currently no protocols available for this strain. | ||
| Donating Investigator: | Dr. Nancy Jenkins | ||
| Key Reference: | Dupuy AJ, Rogers LM, Kim J, Nannapaneni K, Starr TK, Liu P, Largaespada DA, Scheetz TE, Jenkins NA, Copeland NG. A modified sleeping beauty transposon system that can be used to model a wide variety of human cancers in mice. Cancer Res. 2009 Oct 15;69(20):8150-6. Epub 2009 Oct 6. PMID: 19808965 [PubMED Abstract] | ||
| Sponsoring Institute: | National Cancer Institute | ||
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