Study On The Role Of LncRNA3720 During The ZGA Of Goat Embryos In Vitro

Long non-coding RNA(lncRNA)are transcripts that are more than 200 nucleotides in length and are not involved in protein coding,and are also epigenetic regulators.With the development of high-throughput sequencing technology,more and more lncRNA have been shown to play an important role in biological development,and they can be involved in early embryonic development through epigenetic modifications and transcriptional regulation.During mammalian zygotic genome activation(ZGA),a large number of new proteins are synthesized to replace parental material to regulate early embryonic development,and the initiation of ZGA requires the timely and coordinated expression of thousands of genes.The lncRNA and histone modification-related proteins form an action complex that acts at the target site and induces epigenetic modification of genes at that site,regulating the expression of downstream genes,which is essential for embryonic development.During somatic cell nuclear transfer(SCNT)embryo development,ZGA abnormalities may lead to developmental arrest of nuclear transfer embryos at the ZGA stage.The results of numerous studies have resolved the functional mechanisms of lncRNA involvement in mammalian embryonic development,but most of the studies were conducted in mice,and the interspecies conservation of lncRNA is relatively poor,and the mechanisms of action in different mammals are not exactly the same.However,there are few studies on the function of lncRNAs in the process of ZGA in goats.As an important domestic animal,goats have great economic value,both for providing meat and dairy products and as a carrier of mammary gland bioreactors to produce medicinal proteins.Therefore,it is important to investigate the mechanism of lncRNA regulation of early embryonic development in goats and to find ways to improve the efficiency of somatic cell nuclear transfer for the development of animal husbandry and biomedicine.This study explored the role of lncRNA3720 in the goat embryonic ZGA period in vitro based on the previous research of the group.The main contents and results are as follows:1.By interfering with the expression of lncRNA3720 in in vitro fertilized embryos,the expression levels of key congenic genes EIF1 AX and ZSCAN4 were found to be significantly down-regulated(p<0.05)with both potential target genes H2 A type 2-C and H2 B type 1,while microinjection of their in vitro transcribed m RNAs compensated for their expression,and the expression of key congenic genes EIF1 AX and ZSCAN4 were significantly up-regulated with the expression levels of potential target genes H2 A type 2-C and H2 B type 1(p<0.05).2.The nucleoplasmic segregation assay determined that lncRNA3720 was mainly expressed in the nucleus.Using the SCNT technique to produce goat cloned embryos,m RNA transcribed in vitro from lncRNA3720 was microinjected into the cloned embryos,and the expression levels of key congenic genes EIF1 AX and ZSCAN4 were found to be significantly increased(p<0.05),and the expression levels of potential target genes H2 A type 2-C and H2 B type 1 were also significantly increased(p<0.05).By CDX2 staining of cloned blastocysts,it was found that the total number of blastocyst cells and the number of trophoblast cells in the test group were significantly higher than those in the control group(p<0.01),while the number of apoptotic cells in the test group was found to be significantly smaller than that in the control group by TUNEL staining(p<0.05).In summary,interference with lncRNA3720 expression inhibits early embryonic development during goat ZGA,and microinjection of m RNA transcribed from lncRNA3720 in vitro improves the in vitro development of cloned embryos during the development of goat somatic cell nuclear transfer embryos.lncRNA3720 may enhance the in vitro development of cloned embryos by interacting with H2 A type 2-C and H2 B type 1 lncRNA3720 may regulate the reprogramming of goat somatic cell nuclear transfer embryos through interaction with H2 A type 2-C and H2 B type 1,thereby improving cloning efficiency.This study identifies a novel transcript affecting ZGA in goats,which enriches the study of lncRNA regulation of early embryonic development and provides theoretical guidance for improving the development of cloned embryos.