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Improvement and applications of adeno-associated virus 2 (AAV-2) system for efficient gene transfer and expression in penaeid shrimp cells

文献类型： 外文期刊

作者： Tao, Yiwen 1 ; Zhang, Qingli 4 ; Guo, Huarong 1 ;

作者机构： 1.Ocean Univ China, Minist Educ, Key Lab Marine Genet & Breeding, Qingdao 266003, Peoples R China

2.Ocean Univ China, Coll Marine Life Sci, Qingdao 266003, Peoples R China

3.Ocean Univ China, Inst Evolut & Marine Biodivers, Qingdao 266003, Peoples R China

4.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Qingdao 266071, Peoples R China

关键词： Shrimp cells; Adeno-associated virus 2 (AAV-2); Gene transfer; WSSV Tegument protein VP26; IHHNV P2 promoter

期刊名称：AQUACULTURE REPORTS （ 影响因子：3.7； 五年影响因子：3.8 ）

ISSN： 2352-5134

年卷期： 2023 年 32 卷

页码：

收录情况： SCI

摘要： Adeno-associated virus 2 (AAV-2)-mediated expression system is a well-developed and widely used gene transfer and expression tool in mammalian cells, but no attempt has been done to examine its infection and expression efficiency in shrimp cells. In this study, using eGFP as a reporter gene, we found that unmodified AAV-2 could infect primarily cultured shrimp hemolymph cells, but with an extremely low infection efficiency of up to 0.0067%. However, all the improvements of AAV-2 expression system tried in this study including the insertion of shrimp virus (IHHNV)-sourced P2 promoter into the AAV-2 vector (i.e. pAAV-CMV-P2-eGFP) and the inclusion of shrimp virus (IHHNV)-sourced nucleocapsid protein (IHCP) into or association of shrimp virus (WSSV)sourced tegument protein (VP26) with the capsid of the improved AAV-2 s, respectively, could significantly increase the infection and expression efficiencies of this expression system in the primarily cultured shrimp hemolymph cells in comparison with wild-type AAV-2. Moreover, from the viewpoint of improving the infection and expression efficiency, the insertion of P2 promoter did best, followed by the association of WSSV tegument protein of VP26, and the last was the inclusion of IHHNV nucleocapsid protein of IHCP. In conclusion, an improved AAV-2 expression system which consists of four expression plasmids of pAAV-CMV-P2-eGFP, pAAVRC, pHelper and pcDNA3.1-VP26 and one packaging cell line of HEK293T has been successfully developed in this study. This improved AAV-2 expression system will provide us a useful tool for efficient gene transfer and expression in shrimp cells.

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