Summary | Mutation origin | Mutation description | Find Mice (IMSR) | Notes | References

Symbol:	Col1a1tm7(tetO-Pou5f1,-Klf4,-Sox2,-mCherry)Hoch
Name:	collagen, type I, alpha 1; targeted mutation 7, Konrad Hochedlinger
MGI ID:	MGI:6392017
Synonyms:	tetOP-OKSmC
Gene:	Col1a1  Location: Chr11:94827050-94843868 bp, + strand  Genetic Position: Chr11, 59.01 cM, cytoband D
Alliance:	Col1a1tm7(tetO-Pou5f1,-Klf4,-Sox2,-mCherry)Hoch page

Germline Transmission:	Earliest citation of germline transmission: J:284696
Parent Cell Line:	KH2 (ES Cell)
Strain of Origin:	(C57BL/6 x 129S4/SvJae)F1

Allele Type:    Targeted (Inducible, Inserted expressed sequence, Reporter) Inducer:    doxycycline/tetracycline Mutation:    Insertion   Col1a1tm7(tetO-Pou5f1,-Klf4,-Sox2,-mCherry)Hoch expresses 3 genes Knock-in expresses: Organism Expressed Gene Note mouse Klf4 (MGI:1342287) mouse Pou5f1 (MGI:101893) mouse Sox2 (MGI:98364)   Mutation details: A targeting vector was designed to insert a FRT-flanked PGK-Neo-pA into the 3' UTR of the Col1a1 locus. This construct was electroporated into (C57BL/6 x 129S4Sv/Jae)F1-derived V6.5 embryonic stem (ES) cells. Correctly targeted ES cells were identified. One of these ES cells clones, C10, was re-targeted to insert an optimized form of reverse tetracycline controlled transactivator (rtTA-M2) followed by a beta-globin intron, polyA signal and PGK-puromycin cassette, all inserted downstream of the Gt(ROSA)26Sor promoter. Correctly targeted ES cells were identified. One of these ES cell clones, KH2, was selected and the FRT-flanked PGK-Neo-pA in the 3' UTR of the Col1a1 locus was replaced by co-injection of a tetOP-OKSM "flip-in plasmid" and a CAG-Flpe plasmid. The tetOP-OKSmC "flip-in plasmid" contained a splice acceptor-double polyA sequence and the tetracycline responsive element (TRE, tetOP, or tetO) upstream of the three mouse reprogramming genes Pou5f1, Klf4, and Sox2, and an enhanced red fluorescence sequence, mCherry. Internal ribosome entry site (IRES) sequences were placed between the coding sequences for Klf4, Sox2, and mCherry. Expression of Pou5f1, Klf4, and Sox2 was linked by self-cleaving peptides that mediate ribosomal skipping (F2A [from foot-and-mouth disease virus] and E2A [from equine rhinitis A virus], respectively). (J:284696)

Mouse strains and cell lines available from the International Mouse Strain Resource (IMSR)
Carrying this Mutation:	Mouse Strains: 1 strain available      Cell Lines: 0 lines available
Carrying any Col1a1 Mutation:	160 strains or lines available

Original:	J:284696 Bar-Nur O, et al., Small molecules facilitate rapid and synchronous iPSC generation. Nat Methods. 2014 Nov;11(11):1170-6
All:	2 reference(s)