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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/Nci 
 
Release Category (Required for MTA form): C1
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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:
  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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