C57BL/6-Rpl3l^em1Jmol/J

Stock number: 038191
Product name: Rpl3l^-
Strain synonyms: Rpl3l KO
Research areas: Cardiovascular Research, Developmental Biology Research, Internal/Organ Research

Description

Rpl3l^- mice carry a CRISPR/Cas9-made mutation that results in a frameshift and deletion of exon5 of the Rpl3l gene. These mice may be useful for studying development and maintenance of skeletal and cardiac muscle and function.

Detailed description

Rpl3l is one of the four non-canonical riboprotein genes and it encodes the 60S ribosomal protein L3-like protein (RPL3L) that is highly expressed only in cardiac and skeletal muscle. In the mouse, Rpl3l is only expressed postnatally in skeletal muscle and heart myocytes while its paralog, ribosomal protein L3 (Rpl3) is expressed ubiquitously during early development and in all non-myocytes in adulthood. RPL3L has been implicated in the regulation of skeletal muscle mass and cardiac function; and therefore, Rpl3l mutations may be associated with pediatric cardiomyopathies and other heart disorders.

Rpl3l^- mice carry a CRISPR/Cas9-mediated deletion within exon 5 of the Rpl3l gene, resulting in an out of frame null allele. Homozygous mice are viable and fertile, with no overt phenotype. While no changes in cardiac function were observed in the homozygous Rpl3l^- mice, they do demonstrate significantly smaller hearts compared to wildtype controls by 18 months of age. These Rpl3l^- mice may be useful for studying the relationship of Rpl3 and Rpl3l in skeletal and cardiac muscle development and function.

Development

The Rpl3l^- allele was generated using CRISPR/Cas9 endonuclease-genome editing. Two guide RNAs [5'-GTGGGCCTTCTTCTGCCGAA-3' and 5'-TGGCTGAACACGCTGTGTAC-3'] were selected to target exon 5 of the ribosomal protein L3-like (Rpl3l) gene on Chromosome 17. Rpl3l transcript Rpl3l-variant 1 (NM_001163945) was used as reference for the exon number and the guide sequences. These guide sequences and the Cas9 nuclease mRNA were introduced into the cytoplasm of zygotes derived from C57BL/6NTac mice with well recognized pronuclei. Correctly targeted pups were identified by PCR and Sanger sequencing. The resulting Rpl3l^- heterozygous mice were intercrossed for colony maintenance by the donating laboratory (see SNP note below). Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes (Stock No. 005304).

A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the males sent to The Jackson Laboratory Repository. One of the 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.

Allele

Symbol: Rpl3l^em1Jmol
Name: endonuclease-mediated mutation 1, Jeffery D Molkentin
MGI accession ID: MGI:7434982
Generation method: Endonuclease-mediated
Attributes: Null/Knockout
Marker symbol: Rpl3l
Marker name: ribosomal protein L3-like
Marker synonyms: 1110057H16Rik; CMD2D
Chromosome: 17
Strain of origin: C57BL/6NTac
Molecular note: CRISPR/Cas9 technology using sgRNAs 5’-GTGGGCCTTCTTCTGCCGAA-3’ and 5’-TGGCTGAACACGCTGTGTAC-3’generated a frame-shift and subsequent deletion mutation in exon 5. Western blot analysis confirmed absence of protein in heart ventricle.