Establishment of cetp-/- model and liver transcriptomics study in zebrafish

Abstract

Abstract: Objective To investigate the transcriptomic changes in atherosclerosis after cetp knockout of zebrafish. Methods The gene editing technology CRISPR/Cas9 was used to knock out cetp of zebrafish to construct a homozygous mutant model. Total RNA of livers from wild type and cetp^-/- model was extracted by the Trizol method, and gene expression profiles of cetp knockout were observed by transcriptomics. KEGG enrichment analysis was performed on significantly different expressed genes to explore the signal pathways it may affect. Results Homozygous mutant zebrafish that has a frameshift mutation of adding 5 base pairs was created through CRISPR/Cas9 technology. Transcriptomics analysis showed that the gene expression profile of zebrafish liver changed significantly after cetp knockout. A total of 3 808 different expressed genes were screened (P＜0.05), of which 1 918 genes were up-regulated and 1 890 genes were down-regulated. Genes related to lipid metabolism such as abca1 and abcg2a were significantly up-regulated and lipg were significantly down-regulated. Genes related to autophagy signaling pathways such as atg13 were significantly up-regulated, and down-regulated genes were enriched in related pathways such as DNA and RNA synthesis and the tricarboxylic acid cycle. Conclusions A zebrafish model with cetp knockout is established to study the mechanism of cholesteryl ester transfer protein (CETP) in atherosclerotic diseases. Although increased high-density lipoprotein cholesterol due to the inhibition of CETP prevents atherosclero-sis, inflammation is activated after cetp knockout in zebrafish and causes atherosclerosis. It suggests that CETP has a dual regulation mechanism in the development of atherosclerosis.

Key words: cholesteryl ester transfer protein, atherosclerotic cardiovascular disease, CRISPR/Cas9 system, zebrafish, RNA-seq

CLC Number:

R541.4

FENG Dong-ping, SHANG Han-qiao, YANG Hang, ZHANG Hu-jun, ZHANG Ting, YANG Meng-xi, TU Qiang, REN Jing-yi. Establishment of cetp^-/- model and liver transcriptomics study in zebrafish[J]. Basic & Clinical Medicine, 2020, 40(7): 940-947.

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Establishment of cetp^-/- model and liver transcriptomics study in zebrafish

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