Autism spectrum disorders (ASD) is a group of early onset heterogenous neurological disorders with strong genetic predisposition. Due to very high genetic and phenotypic variability and combinations of factors contributing to disease onset, elucidating the mechanism of the disorder is in most cases difficult. Advances in human genetics and functional genomics have brought to light many potential candidate genes, whose roles are yet to be determined. We investigated an ASD candidate gene astrotactin 2 (ASTN2) as a potential risk gene and based on the literature review described its role in cerebellar neural migration as a possible contributing factor for ASD. We tested the potential of CRISPR/Cas9 technology for development of a mouse model of ASD, with which we could in future conduct functional in vivo analyses to determine the associations between molecular, physiological, functional and behavioural factors. This study consisted of preliminary molecular experiments in vitro and in vivo, to determine the feasibility of using this approach to generate a murine Astn2 gene knockout model. We chose the mouse Astn2 exon 2 as our target site for sgRNA and Cas9 cleavage. We confirmed the efficiency of this sgRNA for gene editing using in vitro DNA cleavage assay and GFP-based detection system in the HEK293 cell line by employing the pCAG-EGxxFP plasmid. We concluded that the selected sgRNA is robust and effective for use in further functional experiments in vitro and in vivo.
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