SmsG56S CRISPR/Cas9-generated mutant of the Sms gene carries the G56S point mutation. This strain may be useful in studies related to Snyder-Robinson mental retardation syndrome.
Cathleen Lutz, The Jackson Laboratory
CRISPR/cas9 endonuclease mediated genome editing of the X-linked Sms, spermine synthase, gene was used to introduce the G56S (Glycine at amino acid 56 to a Serine) and an EcoRI site. The targeted Sms gene encodes an enzyme involved in biosynthesis of polyamines. Mutations in this gene have been associated with X-linked Snyder-Robinson mental retardation syndrome. Hemizygous males are infertile and approximately half die by weaning. Heterozygous females are viable and fertile. As the mice are characterized, we will modify the strain description and add phenotype data.
The SmsG56S allele was created by Dr. Cathleen Lutz (The Jackson Laboratory) using CRISPR/cas9 endonuclease-mediated genome editing in C57BL/6J mouse zygotes (Stock No. 000664).
Specifically, the spermine synthase locus (Sms) on chromosome X was targeted with single guide RNAs designed to substitute the GGC wild type glycine56 codon with a TCC Serine56 codon (G56S). The sgRNAs, an oligonucleotide sequence containing the G56S mutation, and Cas9 endonuclease were electroporated into the cytoplasm of single cell C57BL/6J-derived zygotes containing well recognized pronuclei. Embryos were transferred to pseudopregnant females, and correctly targeted pups (identified by DNA sequencing) were bred to C57BL/6J inbred mice for germline transmission. Mice derived from founder 5525 (GET2418) were further bred to C57BL/6J inbred mice for 2 generations to develop the colony. As a consequence of these nucleotide changes the mutant SmsG56S allele contains an EcoRI restriction enzyme site that is absent in the wild type Sms allele.
|Allele Name||endonuclease-mediated mutation 2, Cathy Lutz|
|Allele Type||Endonuclease-mediated (Not Applicable)|
|Gene Symbol and Name||Sms, spermine synthase|
|Strain of Origin||C57BL/6J|
|Molecular Note||CRISPR/Cas9 genome editing is used to substitute the GGC wild type glycine56 codon with a TCC Serine56 codon (G56S). The mutation introduces a EcoRI restriction enzyme site.|
When maintaining a live colony, these mice can be bred as heterozygous females to C57BL/6J (Stock# 000664) or wildtype male littermates (the gene is X linked). Hemizygous males are infertile. As the mice are characterized, we will modify the strain description if necessary and add data related to viability and fertility.
When using the SmsG56S mouse strain in a publication, please cite the originating article(s) and include JAX stock #031170 in your Materials and Methods section.