Small RNA interfering effects in chicken embryonic fibroblasts using two sites 8664/p201 and 2122/pll3.7 of chicken s1p1/edg 1 gene
1 Department of Biology, Laboratory of Biochemistry, Faculty of Sciences, Gamal Abdel Nasser University of Conakry (UGANC), BP: 1147 Conakry, Republic of Guinea.
2 School of Life Science, Laboratory of biochemistry, University of Xiamen, Fujian, People's Republic of China. PO Box: 361005.
Research Article
World Journal of Advanced Research and Reviews, 2022, 16(03), 116-124
Article DOI: 10.30574/wjarr.2022.16.3.1320
Publication history:
Received on 20 October 2022; revised on 29 November 2022; accepted on 02 December 2022
Abstract:
Introduction: Currently, short interfering RNA (siRNA) is an experimental tool in molecular biology to study gene function in cell culture and in vivo in model of organisms (mouse and chicken embryos, etc.)
Method: Lentivirus production technique by human embryonic kidney 293T cell lines (HEK293T) with two (2) sites 8664/P201 and 2122/Pll.3.7 of the sphingosine 1 phosphate receptor gene and endothelial differentiation 1 gene (S1P1/ Chicken EDG1) was used to show the effects of small interfering RNA in chicken embryonic fibroblasts during embryogenesis. After preparation of plasmids 8664/P201 and 2122/Pll3.7, lentivirus packaging, transfection with 2xHBS, virus titration and chicken egg preparation, microinjection of lentivirus was performed in the embryos chicken.
Results: The results obtained after the microinjection of lentivirus into chicken embryos from the second day to the fourth day of incubation showed the effects of small interfering RNA in the embryonic development of chicken materialized by bleeding in the yolk sac of embryos therefore a decrease in gene function.
Conclusion: Previous studies have already proven that expression of small interfering RNA decreases in embryos of certain vertebrates such as mammals.
Keywords:
Lentivirus; HEK293T; Small interfering RNA; Embryonic fibroblasts; Micro injection; Virus titration
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