Developing CRISPR Strategies to Generate Stable Cell Lines that Expressed GABRA1 Subunit for alzheimer Disease Models

Abstract

GABA or gamma-aminobutyric acid is the major neurotransmitter that acts as inhibitors in the central nervous system. It is synthesized by the GAD or glutamic acid decarboxylase and transported into the synaptic system. GABA receptors are a protein consisting of at least 20 different subunits and considered as a complex molecule in the human brain. Specifically, the α1 subunit is abundantly expressed in the stratum radiatum of the CA1 region as it occupies 60% of the total subunit present in the brain. It is known that in Alzheimer Disease patients, the GABA A α1 receptor undergoes some alteration, resulting in many harm risks such as losing the binding site for important drugs as well as the binding of GABA itself which plays a crucial role in preserving memories. The new technology of CRISPR-Cas9 gene editing is expected to be a promising research for Alzheimer Disease models as the process is low on the cost, straightforward, and precise. The model is also chosen because in-vitro model of AD is not yet available. Thus, this in-silico study opens a research area on in-vitro models of AD especially for developing stable cell lines that can express the GABRA1 subunit. The procedure was done by using CRISPR-Cas9 genome editing technology where knock-in strategies to initiate homology directed repair is focused. Thus, the project expects that these procedures can be implemented by hands-on experiments as promising research for in-vitro Alzheimer ’s Disease models in the future.