Screening of Anti-atherosclerosis Drugs Based on the Apolipoprotein apoeb Gene Knockout Zebrafish Model
摘要
Currently, researchers predominantly employ high-cholesterol diets to induce atherosclerosis-like features or perform microinjection and strain establishment procedures targeting individual novel genes to modify animal models. However, it is important to note that while current gene editing technologies simplify model construction, they still present challenges such as high operational redundancy, genetic limitations, and insufficient comprehensiveness in single-strain studies.
ObjectiveWe aim to establish a stable, genetically transmitted apoeb knockout zebrafish strain using CRISPR/Cas9 technology and multiline transgenic zebrafish. This will create a stable, multiline apoeb knockout zebrafish atherosclerosis model platform for drug screening, meeting research demands for atherosclerotic disease studies.
MethodsThis study first employed CRISPR/Cas9 technology to construct a stable, multi-strain zebrafish apoeb mutation model, validated by DNA extraction from tail fin clips. Following successful model establishment, zebrafish were fed different diets. observing characteristics such as Oil Red O staining, immune cell fluorescence reactions, and blood cell aggregation to establish an atherosclerosis (AS) model platform. Subsequently, zebrafish embryos from this platform underwent drug bath treatment, with the same indicators observed. Changes in biochemical and pathological indicators before and after treatment were compared to evaluate the platform's efficacy.
ConclusionThis study successfully established a stable, multi-strain apoeb gene mutation zebrafish atherosclerosis model screening platform. It demonstrated the feasibility of rapid, convenient, and high-throughput drug screening using this platform.