| Primary Identifier | MGI:6113939 | Allele Type | Endonuclease-mediated |
| Attribute String | Modified isoform(s) | Gene | Cdkn2a |
| Strain of Origin | C57BL/6J x C57BL/6N | Is Recombinase | false |
| Is Wild Type | false |
| molecularNote | For CRSIPR/Cas9 mutagenesis, C57BL/6 ES cells were transfected with an all-in-one construct expressing Cas9 and an sgRNA targeting the internal Arf translation initiation codon at position 45 (shown in capital letters; 5'-ctttcgtgaacATGttgttg-3'), and a 127-base single-strand oligodeoxynucleotide (ssODN) template. The template contains, between homology arms, 5'-ctttcgtgaacGCCCtgttgCGC-3'. The template directs the mutation of the downstream ATG codon to GCC (p.Met45Ala), the disruption of the sgRNA-targeting site through a T>C substitution, and the convertion of the 3' PAM sequence from AGG to CGC (all mutations shown in capital letters in the DNA sequence above). The latter two mutations do not affect the sequence of the encoded peptide. |
