Epigallocatechin gallate attenuates neurocognitive impairment in CRISPR-CAS9-induced knockout of DYRK1A gene in zebrafish model

Abstract

Down Syndrome (DS) is the most widespread genetic disorder and a leading cause of mental retardation. It is assumed that the difference in DYRK1A gene expression with Trisomy 21 significantly contributes to several DS traits, including craniofacial or neurological malformation/deformity. Preparation of Epigallocatechin Gallate (EGCG) enriched fraction of Camellia sinensis and identification of compounds by GC–MS chromatogram analysis to identify the presence of bioactive compounds. Determination of total phenolics, flavonoids, DPPH assay, and catalase activity was carried out to quantify its presence. Microinjection during the embryonic stage was performed to observe GFP mCherry transcription through fluorescence imaging. T maze experimentation for various parameters with latency time was observed to test differences between each group. Brain histology was performed to observe neuronal changes in the brain. The total phenolic and flavonoid content shows anti-oxidative potency in EGCG hydroalcoholic extract. After the embryos were microinjected with gene editing materials, the transcription was cumulative with the success of gene editing. During the later phase of development, the knocked fishes had developmental errors in the craniofacial morphology. Morphological visualization reveals a maldevelopment in the gene knockout fish. T maze shows that each category's latency decreases with the increasing dose. EGCG has the potential to ameliorate cognitive impairment in CRISPR CAS9-induced DYRK1A gene knockout zebrafish.