C57BL/6-Ryr1^tm2.1Mfgo/J

Stock number: 035530
Product name: RC-ΔL , Ryr1^{S1669C/L1716Del}
Research areas: Developmental Biology Research, Neurobiology Research

Description

RC-ΔL knock-in mice carry S1669C (serine to cysteine) and L1716Del mutations in the Ryr1 (ryanodine receptor 1, skeletal muscle) gene. Mutations in Ryr1 are associated with RYR1 related myopathies (RYR1 RM). When combined with a mouse carrying the Ryr1^TM allele (Stock No. 033336), progeny develop a severe form of RYR-1 myopathy.

Detailed description

RC-ΔL knock-in mice carry S1669C (serine to cysteine) and L1716Del mutations in the Ryr1 (ryanodine receptor 1, skeletal muscle) gene. Mutations in Ryr1 are associated with RYR1 related myopathies (RYR1 RM). Ryr1 encodes a large homotetrameric Ca2+ release channel located in the sarcoplasmic reticulum that is required for excitation-contraction coupling. Heterozygous mice are viable and fertile with no myopathy phenotype; the mutation is neonatal lethal in the homozygous state.

When combined with a mouse carrying the Ryr1^TM (T4709M) allele (Stock No. 033336), progeny heterozygous for both mutations demonstrate muscle weakness, reduced muscle mass, reduced RYR1 protein expression, kyphosis, malocclusion, and penile prolapse. These mice may be useful for studying congenital myopathies.

Development

The RC-ΔL knock-in allele was created at Ingenious Targeting Laboratory at the direction of the RYR-1 Foundation. A targeting vector was designed to insert an FRT-flanked neomycin-resistance gene upstream of exon 34 and an amino acid substitution at position 1669 altering serine to cysteine (S1669C; TCG to TGC) and a Leu1716 deletion (CTC) into exon 34 of the Ryr1 (ryanodine receptor 1, skeletal muscle) gene on chromosome 7. A silent mutation Leu1715 (CTC to CTG) was introduced to create a Bcl1 enzyme site. The construct was electroporated into C57BL/6-derived iTL BF1 embryonic stem (ES) cells (C57BL/6 FLP), which express a FLP recombinase vector to remove the selection cassette. Correctly targeted ES cells were injected into BALB/c blastocysts. The resulting chimeric animals were bred to C57BL/6J wildtype mice. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Ryr1^tm2.1Mfgo
Name: targeted mutation 2.1, Michael Goldberg
MGI accession ID: MGI:6450020
Generation method: Targeted
Attributes: Humanized sequence
Synonyms: RC-deltaL
Marker symbol: Ryr1
Marker name: ryanodine receptor 1, skeletal muscle
Marker synonyms: calcium release channel isoform 1; CCO; CMYO1A; CMYO1B; CMYP1A; CMYP1B; KDS; MHS; MHS1; PPP1R137; RYDR; Ryr; RYR-1; Ryr1l; SKRR
Chromosome: 7
Strain of origin: C57BL/6NTac
Molecular note: The targeting vector is designed to insert an FRT-flanked neomycin resistance gene upstream of exon 34, an amino acid substitution at position 1669 altering serine to cysteine (S1669C; TCG to TGC) and a Leu1716 deletion (CTC) into exon 34. A silent mutation Leu1715 (CTC to CTG) was introduced to create a Bcl1 enzyme site. The neo cassette was deleted by transient transfection of an expression vector of flp recombinase in the ES cells.