ApoE4 allele specific knockout using a synthetic Cas9 variant as a potential gene therapy approach for Alzheimer's disease

ConclusionMethods:The two alleles of ApoE (E3 and E4) differ in one nucleotide. Based on this single-nucleotide polymorphism (SNP), we designed a novel “clustered regularly interspaced short palindromic repeats” (CRISPR) based system that specifically recognizes, binds and targets the ApoE4 allele for knockout by using the VRER CRISPR associated protein 9 (Cas9) variant.Results and conclusionApplication of this approach in mice astrocytes expressing the human ApoE3 or ApoE4 resulted in a significant decrease of ApoE4 protein levels (-70%) without any significant changes in ApoE3 levels. The cell-culture experiments provide a proof of concept that indeed our CRISPR based system can be used to specifically knockout ApoE4 without affecting ApoE3. These experiments are currently being extended to the in-vivo level. We hope that this study will lead to development of novel gene-therapy applications for AD patients, carriers of the ApoE4 allele in general and ApoE3/4 heterozygous patients in particular.
Source: Cytotherapy - Category: Cytology Source Type: research