The Study On The Effect Of Kmt5b Gene On The Development Of Zebrafish Nervous System

Background With the development of sequencing technology,the research on KMT5 B gene is no longer limited to traditional molecular biological functions,such as cell proliferation and cancer.Through the Next-generation sequencing,more and more mutations of the KMT5 B gene are found in the population.To summarize the clinical manifestations of these mutations,we found that patients with KMT5 B mutations mainly present with neurodevelopmental disorders such as autism and intellectual disability.These suggest us that KMT5 B gene plays a very important role in the development of the nervous system and related diseases,and also provides us with a new direction to study the function of the KMT5 B gene.However,no one has used any suitable animal models and appropriate gene editing techniques to further study the function of KMT5 B.Objective To investigate the effects of the kmt5 b gene on the early development of the nervous system using the zebrafish model.Methods 1.The mutants of zebrafish kmt5 b gene were constructed by CRISPR/Cas9 technology,and functional deletion mutants were screened.The effects of these mutations on zebrafish embryo morphology and development process were observed.2.To examine the spatiotemporal expression profile of the zebrafish kmt5 b gene during early embryo development.3.The RNA-seq technique was used to study the changes in the transcriptional level of the zebrafish kmt5 b mutant.Results 1.We successfully obtained the homozygous mutant of the zebrafish kmt5 b gene using CRISPR/Cas9 technology,and found that brain of the mutant has a delayed development and growth retardation.The whole in situ hybridization of zebrafish embryos showed that the kmt5 b gene was widely expressed in early embryos,and it was mainly concentrated in the brain after 48 h.The q PCR results showed that the kmt5 b gene was abundantly expressed in the early developmental stage,and its expression level became lower and constant as the embryo development progressed.3.RNA-seq sequencing results showed that there were a large number of differentially expressed genes between the kmt5b mutant and the wild-type zebrafish,and there were differences in the functions related to the nervous system.Conclusion Through the observation of the spatiotemporal expression profile of the kmt5 b gene during the early development of zebrafish embryos,the measurement of mutant phenotypes,and the analysis of RNA-seq sequencing results,we believe that the mutation of kmt5 b gene has an impact on the development of the nervous system.