Igs2^{tm1(CAG-mCherry,-dcas9/KRAB)Mtm}
Targeted Allele Detail

Summary

• Symbol: Igs2^{tm1(CAG-mCherry,-dcas9/KRAB)Mtm}
• Name: intergenic site 2; targeted mutation 1, Micheal McManus
• MGI ID: MGI:6441034
• Synonyms: H11^dCas9-KRAB, Igs2^{tm1(CAG-mCherry,-cas9/ZNF10*)Mtm}
• Gene: Igs2 Location: Chr11:3195460-3195497 bp Genetic Position: Chr11, Syntenic
• Alliance: Igs2^{tm1(CAG-mCherry,-dcas9/KRAB)Mtm} page

Mutation origin

• Germline Transmission: Earliest citation of germline transmission: J:278806
• Parent Cell Line: Other (see notes) (ES Cell)
• Strain of Origin: STOCK Igs2^tm3.1Luo/Luo

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: 69 strains or lines available

Notes

This allele was produced in zygotes collected from mice carrying the Igs2^tm3.1Luo allele, which was generated in (129X1/SvJ x 129S1/Sv)F1-Kitl⁺-derived R1 ES cells on a mixed background with 129S1/SvJ, 129X1/SvJ, C57BL/6J, DBA/2J, and FVB/N.

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: 8 reference(s)