Related Experiment Video
Updated: Jan 16, 2026
07:31
Efficient PAM-Less Base Editing for Zebrafish Modeling of Human Genetic Disease with zSpRY-ABE8e
Published on: February 17, 2023
1.6K
Construction and phenotypic analysis of p2rx2 knockout zebrafish lines
Yong Zhang1,2, Qingying Shi3,4,5, Hao Xie3,4,5
1Department of Otolaryngology-Head and Neck Surgery, Second Xiangya Hospital, Central South University, Changsha 410011. 765415941@qq.com.
Summary
We created a zebrafish model with a P2RX2 gene knockout, revealing its crucial role in auditory hair cell function and hearing. This model aids in understanding P2RX2-related hearing loss and developing gene therapies.
Area of Science:
- Genetics and Molecular Biology
- Neuroscience
- Otolaryngology
Background:
- The purinergic receptor P2X2 (P2RX2) is an ATP-gated ion channel crucial for cellular functions.
- Loss-of-function mutations in P2RX2 cause autosomal dominant nonsyndromic deafness 41 (DFNA41), characterized by hearing loss and noise susceptibility.
- Zebrafish are an ideal model organism for studying human diseases due to genetic conservation and developmental transparency.
Purpose of the Study:
- To generate a p2rx2 knockout zebrafish model using CRISPR/Cas9.
- To investigate the impact of p2rx2 deficiency on the auditory system.
- To provide a basis for understanding P2RX2-related hearing loss and developing gene therapy strategies.
Main Methods:
- CRISPR/Cas9 gene editing was employed to target the zebrafish p2rx2 gene.
- Zebrafish embryos were injected, and edited F0, F1, and F2 generations were screened and genotyped.
- Hair cell morphology and density were assessed using YO-PRO-1 staining, and auditory function was evaluated via auditory evoked potential (AEP) testing.
Main Results:
- A stable p2rx2 knockout zebrafish line was successfully established with a 66 bp insertion causing premature protein termination.
- Mutant zebrafish exhibited reduced hair cell density in the otolith region but not in lateral line neuromasts.
- Homozygous mutants showed significantly elevated auditory thresholds across tested frequencies, indicating impaired hearing sensitivity.
Conclusions:
- The generated p2rx2 knockout zebrafish model demonstrates the gene's critical role in auditory hair cell function and hearing perception.
- p2rx2 deficiency leads to otolith hair cell defects and hearing impairment, suggesting tissue-specific functions.
- This zebrafish model is valuable for studying P2RX2-related hearing loss mechanisms and for screening potential otoprotective drugs and gene therapies.
