Gt(ROSA)26Sor^{em3(CAG-Brainbow2.1,-Pik3ca*)Koo}
Endonuclease-mediated Allele Detail

Summary

• Symbol: Gt(ROSA)26Sor^{em3(CAG-Brainbow2.1,-Pik3ca*)Koo}
• Name: gene trap ROSA 26, Philippe Soriano; endonuclease-mediated mutation 3, Bon-Kyoung Koo
• MGI ID: MGI:7259837
• Synonyms: Red2-PIK3CA^H1047R
• Gene: Gt(ROSA)26Sor Location: Chr6:113044389-113054205 bp, - strand Genetic Position: Chr6, 52.73 cM
• Alliance: Gt(ROSA)26Sor^{em3(CAG-Brainbow2.1,-Pik3ca*)Koo} page

Mutation origin

• Germline Transmission: Earliest citation of germline transmission: J:322623
• Parent Cell Line: Other (see notes) (ES Cell)
• Strain of Origin: B6.129P2-Gt(ROSA)26Sor^{tm1(CAG-Brainbow2.1)Cle}/J

Mutation description

• Allele Type: Endonuclease-mediated (Conditional ready, Inserted expressed sequence, No functional change, Reporter)
• Mutation: Insertion
• Mutation details: CRISPR-targeting inserted the Red2Onco construct, containing PIK3CA^H1047R (addgene 12524), into the tdimer2 reporter within the Gt(ROSA)26Sor^{tm1(CAG-Brainbow2.1)Cle}. The Brainbow2.1 (also known as Confetti) reporter construct is a modification of the Brainbow reporter with a strong CAGG promoter, a loxP site, a PGK-Neo-pA cassette (serving as a transcriptional roadblock), and coding region composed of two adjacent floxed head-to-tail tandem dimers. The first head-to-tail dimer contains a loxP site and humanized Renilla GFP (hrGFPII; with nuclear localization signal plus polyA sequence) in forward orientation and a loxP site and monomeric EYFP (mYFPA206K plus polyA sequence) in reverse orientation. The second head-to-tail dimer contains a loxP site and tdimer2 RFP plus polyA sequence in forward orientation, and a loxP site and mCerulean CFP (with membrane tethering palmitoylation sequence plus polyA sequence) in reverse orientation. A single frt site is located at the 3' end of the reporter construct. (J:101977, J:322623)

Find Mice (IMSR)

• Mouse strains and cell lines available from the International Mouse Strain Resource (IMSR)
• Carrying this Mutation: Mouse Strains: 0 strains available Cell Lines: 0 lines available
• Carrying any Gt(ROSA)26Sor Mutation: 1205 strains or lines available

Notes

ES cells made from B6.129P2-Gt(ROSA)26Sor/J mice are used. Last Name : Davis First Name : Crystal Email Address : crystal.davis@jax.org Email Address (repeat) : crystal.davis@jax.org -------------------------------------------------- Are your data published? : yes You would prefer that your data : be public at the MGI website now References : (a href="https://www.informatics.jax.org/allele/MGI:7259837")https://www.informatics.jax.org/allele/MGI:7259837(/a) -------------------------------------------------- Suggested strain name : B6(129P2)-Gt(ROSA)26Sor(em3(CAG-Brainbow2.1,-Pik3ca*)Koo)/J Repository of strain : JAX Repository ID or MGI ID of strain : 041597 Strain categories : mutant strain -------------------------------------------------- Additional Comments or Information about your data : The Red2Pik3ca(H1047R) knock-in reporter allele was generated, through CRISPR/Cas9-assisted homologous recombination, by introducing the Red2Onco construct, containing a 2A peptide sequence and mutant phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (Pik3ca) sequence with the H1047R mutation, downstream of the tdimer2 reporter sequence within the R26R-Brainbow2.1 mouse line (Stock No. (a href="https://www.jax.org/strain/017492")017492(/a)). The R26R-Brainbow2.1, also referred to as R26R-Confetti, mouse line was originally generated with a targeting vector containing (from 5' to 3') a strong CAGG promoter (CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter), a loxP site, a PGK-Neo(r)-pA cassette (serving as a transcriptional roadblock), and the Brainbow 2.1 construct (described in greater detail below). This entire construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus via e lectroporation into 129P2/OlaHsd-derived IB10/E14IB10 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric males were bred with C57BL/6 females to generate the R26R-Confetti colony. This line was further backcrossed to generate a C57BL/6J-congenic strain, offered as Stock No. 017492. The Brainbow2.1 construct was designed with four fluorescent protein sequences uniquely positioned in a tandem fashion and delimited by loxP sites in opposite orientation. Specifically, the Brainbow 2.1 coding region is composed of two adjacent floxed head-to-tail tandem dimers. The first head-to-tail dimer contains a loxP site and humanized Renilla GFP (hrGFPII; with nuclear localization signal plus polyA sequence) in forward orientation, and a loxP site and monomeric EYFP (mYFP(A206K) plus polyA sequence) in reverse orientation. The second head-to-tail dimer contains a loxP site and tdimer2 RFP plus polyA sequence in forward orientation, and a l oxP site and mCerulean CFP (with membrane tethering palmitoylation sequence plus polyA sequence) in reverse orientation. A single frt site is located at the 3' end of the Brainbow 2.1 construct. Dr. Bon-Kyoung Koo acquired some R26R-Brainbow2.1 mice and modified these to create the Red2Pik3ca(H1047R) knock-in reporter mouse line. The Red2Onco construct, containing a mutant phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (Pik3ca) sequence with the H1047R mutation (Addgene plasmid #12524), was inserted into the tdimer2 reporter locus within the R26R-Brainbow2.1 allele. The sgRNA, donor plasmid and Cas9 nuclease were introduced into R26R-Brainbow2.1-derived (Stock No. 017492) embryonic stem (ES) cells. Correctly targeted pups were identified by long-range PCR. Correctly targeted ES cell clones were identified by long-range PCR and used to generate chimeric mice. Resulting Red2Pik3ca(H1047R) mice were maintained on a C57BL/6 background by the donating laboratory. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. (a href="https://www.jax.org/strain/000664")000664(/a)) oocytes. -------------------------------------------------- Submission Directory: /export/gondor/submissions/2026/04/14_50359kw4

References

• Original: J:322623 Yum MK, et al., Tracing oncogene-driven remodelling of the intestinal stem cell niche. Nature. 2021;594(7863):442-447
• All: 2 reference(s)