B6;129-Utf1^em1Hoch/J

Stock number: 034362
Product name: Utf1-GFP
Research areas: Cell Biology Research, Research Tools

Description

Utf1-GFP mice have a CRISPR/Cas9-generated E2A-GFP cassette inserted in-frame with the final exon of the undifferentiated embryonic cell transcription factor 1 (Utf1) gene.

Detailed description

CRISPR/cas9 endonuclease mediated genome editing of the undifferentiated embryonic cell transcription factor 1 (Utf1) gene was used to introduce E2A-GFP cassette in-frame with the final exon. Utf1 encodes a leucine zipper-containing transcriptional coactivator expressed in pluripotent embryonic stem (ES) cells including a subset of early spermatogonia. The UTF1 protein functions as a transcriptional repressor and is involved in the differentiation of embryonic carcinoma and ES cells. EGFP expression in these mice is visible by direct fluorescence. Heterozygous mice are viable and fertile. The donating investigator did not test the fertility of homozygous mice.

Development

Plasmids encoding a sgRNA (5'(GACTGATAACAAAGCTTTAT-3')), designed to introduce an E2A-GFP cassette in-frame with the final exon of the undifferentiated embryonic cell transcription factor 1 (Utf1) gene, and the cas9 nuclease were cotransfected into (C57BL/6 x 129S4/SvJae)F1-derived V6.5 embryonic stem (ES) cells. These ES cells had previously been targeted with doxycycline inducible Col1a1-OKSM and Rosa26-rtTA. Correctly targeted ES cells were injected into recipient blastocysts and resulting chimeric males were bred to C57BL/6 females. This line was maintained on a mixed C57B6/129SvJae genetic background. The Col1a1-OKSM and Rosa26-rtTA alleles were bred out to the mice that arrived at The Jackson Laboratory. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes. An ampicillin cassette is still present in this strain from the CRISPR editing, and has been found to be segregating with the CRISPR mutation.

Allele

Symbol: Utf1^em1Hoch
Name: endonuclease-mediated mutation 1, Konrad Hochedlinger
MGI accession ID: MGI:6368147
Generation method: Endonuclease-mediated
Attributes: Reporter
Synonyms: Utf1-GFP
Marker symbol: Utf1
Marker name: undifferentiated embryonic cell transcription factor 1
Chromosome: 7
Strain of origin: (C57BL/6 x 129S4/SvJae)F1
Site of expression: Pluripotent embryonic stem (ES) cells including a subset of early spermatogonia.
Molecular note: Plasmids encoding an sgRNA (5'(GACTGATAACAAAGCTTTAT-3'))are designed to introduce an E2A-GFP cassette in-frame in the final exon of the gene.
General note: These V6.5 ES cells were previously targeted with doxycycline inducible Col1a1^{tm1(tetO-Pou5f1,-Klf4,-Sox2,-Myc)Hoch} and Gt(ROSA)26Sor^{tm1(rtTA*M2)Jae}.