| Primary Identifier | MGI:7662591 | Allele Type | Endonuclease-mediated |
| Attribute String | Inserted expressed sequence | Gene | Gt(ROSA)26Sor |
| Strain of Origin | C57BL/6J | Is Recombinase | false |
| Is Wild Type | false |
| molecularNote | Using CRISPR/Cas9 technology, the intron 1 of the mouse Gt(ROSA)26Sor locus was targeted to insert LoxP-Stop-LoxP (LSL)-codon-optimized enAsCas12a-HF1 cDNA. EnAsCas12a-HF1 is an engineered, high-fidelity version of the Acidaminococcus sp. Cas12a variant (AsCas12a) with substitutions E174R (GAG to AGA)/N282A (AAT to GCC)/S542R (TCT to AGA)/K548R (AAG to AGA), an expanded PAM sequence, and enhanced multiplexed gene editing efficiency. Egl-13, a nuclear localizing signal (NLS) derived from C. elegans, was placed at the N-terminus, and the nucleoplasmin NLS and c-Myc NLS signals were placed on the C-terminus of enAsCas12a-HF1 to maintain a higher concentration of enAsCas12a-HF1 in the nucleus and thus enhance gene editing efficiency. |
