Establishment of a mouse model of mucopolysaccharidosis-plus syndrome using CRISPR-Cas9 technology

Abstract

Mucopolysaccharidosis-plus syndrome (MPSPS) is an autosomal recessive disease caused by a missense variant in the VPS33A gene. The aim of this work is to create a mouse model of MPSPS by introducing the target variant using CRISPR-Cas9 technology. Sperm and oocyte donors were mice of the C57BL/6J strain. The CRISPR-Cas9 system was introduced by electroporation. Litter birth was observed on days 19-21. Validation of successful variant introduction was performed by PCR-RFLP and Sanger sequencing. This resulted in one mouse (male) in which the p.R500W variant in the Vps33a gene was introduced into the genome. After several crosses, heterozygous and homozygous mice were obtained. Homozygous mice were developmentally retarded. This may be due to the influence of the variant on intrauterine development.