Cloning,expression and knockout model construction of a novel lincRNA-Abcc10 gene in northern pike(Esox lucius)
Long non-coding RNA(lncRNAs)are involved in various biological processes,including sex differentiation.However,most studies on lncRNAs are limited to mammals,and a few studies are available on their functions in teleost fish.In this study,the sex-specific gene Nove101784 was identified using 126-day-old gonadal transcriptome data from northern pike(Esox lucius),and the full-length 1328 bp sequence was cloned using RACE.The results of CPC2 and CPAT analyses showed that the gene did not encode proteins.BLAST alignment analysis revealed that it was located on chromosome 9,between the sp2 and Abcc10 genes.Based on its location,it was named lincRNA-Abcc10.Transcriptome analysis was used to analyze the expression profiles of lincRNA-Abcc10 and its adjacent 11 genes in the ovaries of northern pikes at 126,188,and 320 d.The results showed that lincRNA-Abcc10 was downregulated over time,and Mrpl4 and Mrpl45 were downregulated at all time points,similar to the expression pattern of lincRNA-Abcc10.According to the gene annotations for Mrpl4 and Mrpl45,lincRNA-Abcc10 may be involved in the regulation of ion transport,follicular cell development,and mitochondrial activity.Second,the qRT-PCR results of lincRNA-Abcc10 in eight northern pike tissues at three stages of showed that on day 126,the expression of lincRNA-Abcc10 in the female ovary was significantly higher than that in the testis,whereas in other tissues,it was only slightly expressed in the male muscle.On day 188,the expression of lincRNA-Abcc10 in the ovaries was significantly higher than that in the testes.In other tissues,it was slightly expressed in the liver,kidney,and muscle of female fish and in the head kidney and muscle of male fish.At 320 d,the expression of copyright Abcc10 in the ovaries was significantly higher than that in the testes.Other tissues included the brain,intestines,muscles,and kidneys of male fish.According to the comparative analysis of the relative expression in gonads at different developmental stages,the results showed that with the continuous development of the northern pike,the expression of lincRNA-Abcc10 at each stage decreased significantly compared to the previous period.It has also been suggested that it plays an important role in early ovarian differentiation.Additionally,through the isolation of nuclear RNA from 126-day-old northern pike ovaries,qRT-PCR results showed that lincRNA-Abcc10 was distributed in the cytoplasm and might regulate the translation level of adjacent genes in a cis or trans manner.Finally,a lincRNA-Abcc 10 mutation model was established using the CRISPR/Cas9 technology,which laid the foundation for further studies on its gene function.This study provides new insights into the roles of lncRNAs in fish sex differentiation.
long non-coding RNAepigeneticsEsox luciussexual developmentCRISPR/Cas9
国家科技期刊平台
NSTL
万方数据
