Construction method of zebrafish gene mutation model of retinitis pigmentosa type 58

CN122344576APending Publication Date: 2026-07-07INST OF AQUATIC LIFE ACAD SINICA

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Patent Information

Authority / Receiving Office: CN · China
Patent Type: Applications(China)
Current Assignee / Owner: INST OF AQUATIC LIFE ACAD SINICA
Filing Date: 2026-06-01
Publication Date: 2026-07-07

AI Technical Summary

Technical Problem

Existing technologies for constructing animal models of retinal-related diseases suffer from problems such as low targeting accuracy, phenotypic instability caused by chimerism, and complicated and time-consuming subsequent screening and validation processes. They are unable to quickly and stably reproduce the core pathological features of specific retinal degenerative diseases and cannot meet the needs of exploring the deep mechanisms of diseases and high-throughput drug screening.

Method used

Using sgRNA that specifically binds to the znf513a gene, Cas9 mRNA and sgRNA were mixed and introduced into zebrafish single-cell stage fertilized eggs via microinjection for precise gene editing, thus constructing a zebrafish model of retinitis pigmentosa type 58. The method included microinjection buffer and identification primer pairs to ensure targeting and high efficiency.

Benefits of technology

It significantly reduced off-target effects, improved the specificity and reliability of gene editing, stably reproduced pathological features of retinal diseases, provided highly consistent animal models for drug screening, and enhanced the reliability of research and screening.

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Abstract

The application provides a method for constructing a retinal pigment degeneration type 58 zebrafish gene mutation model, and relates to the technical field of genetic engineering. The sgRNA comprises a guide sequence; the guide sequence can specifically recognize and combine with the nucleotide sequence shown as SEQ ID NO. 4 in the znf513a gene. By specific targeting design for specific sequences, the off-target effect of conventional tools is effectively overcome, and the risk of unintended modification is significantly reduced. The precise recognition mechanism not only improves the directional cutting efficiency, but also provides a reliable molecular basis and source guarantee for stably introducing specific mutations and obtaining retinal disease individuals with consistent phenotypes.

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