C57BL/6NJ-Fkrp^em1Lgmd/J

Stock number: 034659
Product name: Fkrp^P448L
Research areas: Neurobiology Research

Description

Fkrp^P448L is a CRISPR/Cas9-generated c.1343 C->T point mutation in exon 3 of the mouse Fkrp (fukutin related protein) gene that results in a P448L (p.Pro448Leu) substitution in the translated peptide. An additional nucleotide change (c.1353 C>A) encodes a silent mutation (P451P, CCC->CCA) and has been created for genotyping purposes. The phenotype has not been characterized, but homozygotes are anticipated to demonstrate features of LGMD2I (limb-girdle muscular dystrophy type 2I).

Detailed description

Fkrp^P448L is a CRISPR/Cas9-generated c.1343 C->T point mutation in the mouse Fkrp (fukutin related protein) gene that results in a P448L (p.Pro448Leu) pathogenic substitution in the translated peptide. The phenotype has not been characterized, but homozygotes are anticipated to demonstrate features of LGMD2I (limb-girdle muscular dystrophy type 2I).

Homozygous viability and fertility have not yet been fully investigated, but some neonatal/postnatal lethality and reduced body size are expected. Preliminary information suggests approximately 30% of homozygotes die at around postnatal day 2 (P2), and if they live beyond that, there are no further viability issues.

Development

The Fkrp^P448L knock-in allele was generated using CRISPR/Cas9 gene editing. A pair of sgRNAs targeting exon 3 of the mouse Fkrp gene, a pair of single-stranded oligodeoxynucleotide (ssODN) templates for homology-directed repair (HDR), and Cas9 mRNA were injected into C57BL/6NJ embryos. This resulted in a c.1343 C->T pathogenic point mutation (p.Pro448Leu, P448L), and a c.1353 C>A silent mutation (P451P, CCC>CCA) created for genotyping purposes. This strain was created and maintained on a C57BL/6NJ (Stock No. 005304) genetic background at The Jackson Laboratory for the LGMD2i Research Fund and the CureLGMD2i Foundation. This is founder line 8373 was backcrossed to C57BL/6NJ for two generations prior to entering the JAX Repository program for distribution.

Allele

Symbol: Fkrp^em1Lgmd
Name: endonuclease-mediated mutation 1, Jean-Pierre Laurent
MGI accession ID: MGI:6386788
Generation method: Endonuclease-mediated
Attributes: Humanized sequence
Synonyms: Fkrp^P448L
Marker symbol: Fkrp
Marker name: fukutin related protein
Marker synonyms: A830029B19Rik; FKTR; LGMD1I; LGMD2I; LGMDR9; MDC1C; MDDGA5; MDDGB5; MDDGC5
Chromosome: 7
Strain of origin: C57BL/6NJ
Molecular note: CRISPR/Cas9 genome editing is used to introduce a c.1343 C-to T point mutation (p.Pro448Leu, P448L), and an additional nucleotide change encoding a silent mutation (P451P, CCC to CCA). The mutation is associated with severe congenital muscular dystrophy (CMD).