Igs2^{tm1(CAG-mCherry,-cas9/ZNF10*)Mtm}
Targeted Allele Detail

Summary

• Symbol: Igs2^{tm1(CAG-mCherry,-cas9/ZNF10*)Mtm}
• Name: intergenic site 2; targeted mutation 1, Micheal McManus
• MGI ID: MGI:6441034
• Synonyms: H11^dCas9-KRAB
• Gene: Igs2 Location: Chr11:3195460-3195497 bp Genetic Position: Chr11, Syntenic
• Alliance: Igs2^{tm1(CAG-mCherry,-cas9/ZNF10*)Mtm} page

Mutation origin

• Germline Transmission: Earliest citation of germline transmission: J:278806
• Parent Cell Line: R1 (ES Cell)
• Strain of Origin: (129X1/SvJ x 129S1/Sv)F1-Kitl⁺

Mutation description

• Allele Type: Targeted (Endonuclease, Reporter)
• Mutation: Insertion
• Mutation details: The targeting construct contains (from 5' to 3') an attB site, a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from pCAGGS), a viral 2A oligopeptide sequence (T2A; mediates ribosomal skipping), the mCherry gene, a viral 2A oligopeptide sequence (F2A; mediates ribosomal skipping), the dCas9/ZNF10* fusion gene followed by a nuclear localization signal (NLS), an SV40 polyA signal and a second attB site. The dCas9/ZFN10* fusion gene sequence (dK) is composed of a human codon-optimized, nuclease-deficient Streptococcus pyogenes Cas9 protein synthetic DNA fused at its C-terminus to the repressive chromatin modifier domain mRNA from the Kruppel associated box (KRAB) domain of the human zinc-finger protein 10 gene (ZNF10). PhiC31 integrase-mediated transgenesis is used to insert the construct into the first and second attP sites of Igs2^tm3.1Luo. The resulting knock-in allele is therefore H11::pCA::puro::T2A::mCherry::F2A::dCas9/KRAB-NLS-SV40pA::attP::frt5. The dCas9/KRAB binds to a specific DNA sequence/locus following introduction of a single guide RNA (sgRNA), and that locus is subsequently silenced via heterochromatin formation and/or DNA methylation induced by the KRAB domain. (J:278806)

Find Mice (IMSR)

• Mouse strains and cell lines available from the International Mouse Strain Resource (IMSR)
• Carrying this Mutation: Mouse Strains: 2 strains available Cell Lines: 0 lines available
• Carrying any Igs2 Mutation: 72 strains or lines available

Notes

TARGATT mice carry the H11 allele ( Igs2^tm3.1Luo) or H11P3; a unique sequence from the promoter of the yeast his3 gene followed by three shortened tandem attP sites and one FRT5 site. One founder mouse carried the correctly targeted transgenic insertion of the construct into the first and second attP sites of the locus.

References

• Original: J:278806 Yoneshiro T, et al., BCAA catabolism in brown fat controls energy homeostasis through SLC25A44. Nature. 2019 Aug;572(7771):614-619
• All: 4 reference(s)