Preliminary Study On The Effects Of YT521-B And DYTHDF On Embryonic Development In Drosophila

The genes that regulate the development of Drosophila embryos have been continuously studied.Recent studies have shown that genes related to m6A can be involved in regulating the development of genes in Drosophila.Studies have shown that m6A reading protein is involved in the regulation of MZT(maternal to zygote transformation)of embryonic development.Find genes that are highly conserved evolutionarily and homologous in Drosophila.YT521-B participates in m RNA splicing after m6A modification;DYTHDF participates in the process that can promote the translation efficiency of m6A modified m RNA.This article focuses on the function of two YTH domain-containing genes,YT521-B and d YTHDF,in embryos of Drosophila.1.Microbial injection of sg RNA into CRISPR/Cas9 Drosophila to obtain two read protein gene knockout mutants in four strains of Drosophila.They are d YTHDF ~1/TM3,d YTHDF ~2/TM3,YT521-B ~1/TM3,YT521-B ~2/TM3.The genetic background was removed by pairwise crosses between the lines to obtain a double mutant Drosophila,YT521-B ~1/YT521-B ~2,d YTHDF ~1/d YTHDF ~2.2.we purified the prokaryotic expressed protein of the partial fragment of the gene and sent it to make antibodies to test whether the prepared gene mutant Drosophila was completely knocked out.The stability of the mutant Drosophila was further verified.3.In order to determine the relationship between the two genes and m6A respectively,EMSA experiments were performed to detect the binding of the m RNA probe of probe A or probe m6A after the YTH domain proteins of the two genes were purified.Through experiments,we see that DYTHDF does not bind probe A and probe m6A.YT521-B can selectively bind to probe m6A.4.the fertility of the mutant fruit fly was determined by counting the embryo survival rate.The results showed that the embryo survival rate of the double mutant fruit fly of the two genes was not affected by the loss of the maternal source,and the survival rate was normal in the case of the absence of the zygotic gene.In summary,this study proved that the homologous genes YT521-B and d YTHDF in Drosophila that have the same YTH domain in mammals cannot affect the embryonic development of Drosophila through m6A methylation.We speculate that in Drosophila there are other genes that control the development of the embryo.