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Strain
B6J.129(B6N)-Gt(ROSA)26Sortm1(CAG-cas9*,-EGFP)Fezh/J

Summary Data Summary
Official Name B6J.129(B6N)-Gt(ROSA)26Sortm1(CAG-cas9*,-EGFP)Fezh/J
Common Name B6J.129(B6N)-Gt(ROSA)26Sortm1(CAG-cas9*,-EGFP)Fezh/J
Description These Rosa26-LSL-Cas9 knockin mice have a floxed-STOP
cassette preventing expression of the downstream bicistronic
sequences (Cas9 and EGFP). Although under control of a CAG
promoter, widespread expression of cas9 and EGFP is
prevented by the STOP cassette. After exposure to Cre
recombinase, expression of cas9 and EGFP is observed.
The Rosa26-LSL-Cas9 targeting vector used was designed with
(from 5' to 3') a 5' homology arm, a ubiquitously expressed
CAG promoter, loxP-flanked 3xSV40 polyA stop cassette, a
3X-FLAG epitope tag, a mammalian codon-optimized cas9 gene
(derived from Streptococcus pyogenes CRISPR associated
protein 9 [SpCas9]) flanked by two nuclear localization
signals, a P2A ribosomal skip cleavage peptide sequence, an
enhanced Green Fluorescent Protein (EGFP) gene, a woodchuck
hepatitis virus post-transcriptional regulatory element,
bovine growth hormone polyA, pPGK-Neo-pA positive selection
cassette and a 3' homology arm. The construct was
electroporated into 129S-derived R1 embryonic stem (ES)
cells. Correctly targeted ES cells were injected into
C57BL/6N blastocysts. The resulting chimeric animals were
tested for germline transmission by crossing to C57BL/6N
mice. The mice were then backcrossed for four generations
onto C57BL/6N.
Development Status Phenotyping ongoing
Creation Method congenic
TypeCount
Investigators 1
Experiments 2


Investigators
NameInstitution
David OlsonUniversity of Michigan-Ann Arbor


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