Primary Identifier | MGI:6886058 | Allele Type | Endonuclease-mediated |
Attribute String | Humanized sequence, Inserted expressed sequence | Gene | Mecp2 |
Strain of Origin | C57BL/6J | Is Recombinase | false |
Is Wild Type | false |
molecularNote | CRISPR/cas9 genome editing used two guides sgRNA-49229 (5'-gattgcatactttgaaaagg-3') and sgRNA-49791 (5'-ccacccttggtgagaaaagc-3') to target the start and end of the humanization region (exon 4) as well as a single strand DNA (ssDNA) donor template that spans the two cut sides. The homology arm length for the ssDNA donor is 300 nt. The insertion includes a 561 bp humanized MECP2 region, and a Rett Syndrome-associated MECP2 R270X mutation (arginine to stop, c.808 C to T, p.Arg 270*) in exon 4. The 561bp humanized MeCP2 region corresponds to amino acid (aa) positions 145-331 aa in the human MeCP2 locus (Note: brain enriched isoform MeCP2-E1 is 498 aa in length). |