Skip to main content

Advertisement

SpringerLink
Log in

The Improvement and Application of Lentivirus-Mediated Gene Transfer and Expression System in Penaeid Shrimp Cells

  • Original Article
  • Published:
Marine Biotechnology Aims and scope Submit manuscript

Abstract

This study first reported the improvement and application of lentivirus-mediated gene transfer and expression system in shrimp cells. After modified by the inclusion of two envelope proteins (VP19 and VP28) of shrimp white spot syndrome virus (WSSV) into the envelope of the packaged lentivirus, and insertion of a truncated promoter of immediate-early gene 1 (Pie1-504) of shrimp WSSV virus into the lentiviral reporter plasmid, the second-generation lentiviral expression system (pLVX-PEF1α-IRES-mCherry, psPAX2, and PMD2.G) was found to behave better in the mitosis-arrested shrimp cells than the similarly modified retrovirus expression system did. Results from the insect sf9 cells indicated that the inclusion of VP19 and VP28 into the envelope of packaged lentiviruses could significantly improve the tropism or infectivity of the modified lentiviruses to insect cells in a cumulative way. Notably, the VP28 contributed about 86% of the total increase of the tropism. In the shrimp primary lymphoid cells infected by modified lentivirus IV with both VP19 and VP28 included, the infection efficiency was up to 11% (non-confocal) and 19% (confocal) and no background fluorescent signal was observed. However, background fluorescent signal was observed in the shrimp primary Oka organ cells although only under a confocal microscope. In the lentivirus IV-infected Oka organ cells, the actual infection efficiencies were calculated up to 8% (non-confocal) and 19% (confocal), significantly higher than those of commercial intact lentivirus I of 0 (non-confocal) and 3% (confocal). The insertion of WSSV promoter (Pie1-504) had interrupted the effective expression of reporter plasmid encoding lentiviral construct of pLVX-PEF1α-Pie1-504-IRES-mCherry in the HEK293T cells, but markedly increased its efficiencies up to 14% (non-confocal) and 26% (confocal) in the Oka organ cells. This improved lentivirus expression system will provide us a useful tool for efficient gene transfer and expression in shrimp cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Bensheng J, Khoo HW (1997) Transient expression of two luciferase reporter gene constructs in developing embryos of Macrobrachium lanchesteri (De Man). Aquac Res 28:183–190

    Article  Google Scholar 

  • Blömer U, Naldini L, Kafri T, Trono D, Verma IM, Gage FH (1997) Highly efficient and sustained gene transfer in adult neurons with a lentivirus vector. J Virol 71(9):6641–6649

    PubMed  PubMed Central  Google Scholar 

  • Bukrinsky MI, Haggerty S, Dempsey MP, Sharova N, Adzhubei A, Spitz L, Lewis P, Goldfard B, Emermon M, Stevenson M (1993) A nuclear localization signal within HIV-1 matrix protein that governs infection of non-dividing cells. Nature 365(6447):666–669

    Article  CAS  Google Scholar 

  • Chen SN, Chi SC, Kou GH, Liao IC (1986) Cell culture from tissues of grass prawn, Penaeus monodon. Fish Pathol 21(3):161–166

    Article  Google Scholar 

  • Chou HY, Huang CY, Wang CH, Chiang HC, Lo CF (1995) Pathogenicity of a baculovirus infection causing white spot syndrome in cultured penaeid shrimp in Taiwan. Dis Aquat Org 23(3):165–173

    Article  Google Scholar 

  • Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D, Naldini L (1998) A third-generation lentivirus vector with a conditional packaging system. J Virol 72(11):8463–8471

    CAS  PubMed  PubMed Central  Google Scholar 

  • Friesen PD (1997) Regulation of baculovirus early gene expression. In: Miller LK (ed) The baculoviruses. Springer Science and Business Media, LLC, New York

    Google Scholar 

  • Gallay P, Swingler S, Aiken C, Trono D (1995) HIV-1 infection of nondividing cells: C-terminal tyrosine phosphorylation of the viral matrix protein is a key regulator. Cell 80(3):379–388

    Article  CAS  Google Scholar 

  • Gallay P, Hope T, Chin D, Trono D (1997) HIV-1 infection of nondividing cells through the recognition of integrase by the importin/karyopherin pathway. Proc Natl Acad Sci 94(18):9825–9830

    Article  CAS  Google Scholar 

  • Gao H, Wang Y, Li N, Peng WP, Sun Y, Tong GZ, Qiu HJ (2007) Efficient gene delivery into mammalian cells mediated by a recombinant baculovirus containing a whispovirus ie1 promoter, a novel shuttle promoter between insect cells and mammalian cells. J Biotechnol 131:138–143

    Article  CAS  Google Scholar 

  • Gasmi M, Glynn J, Jin MJ, Jolly DJ, Yee JK, Chen ST (1999) Requirements for efficient production and transduction of human immunodeficiency virus type 1-based vectors. J Virol 73:1828–1834

    CAS  PubMed  PubMed Central  Google Scholar 

  • Gendreau S, Lardans V, Cadoret JP, Mialhe E (1995) Transient expression of a luciferase reporter gene after ballistic introduction into Artemia franciscana (Crustacea) embryos. Aquaculture 133:199–205

    Article  Google Scholar 

  • Guo Z, Wang Y, Rong X, Liao M, Guo H, Han Q (2014) The primary culture of the cell from lymphoid organ of Metapenaeus ensis and the infected feature with white spot syndrome virus (WSSV). J Fish China 38(4):140–148

    Google Scholar 

  • Han Q, Li P, Lu Q, Guo Z, Guo H (2013) Improved primary cell culture and subculture of lymphoid organs of the greasyback shrimp Metapenaeus ensis. Aquaculture 410-411:101–113

    Article  Google Scholar 

  • Han Q, Dong D, Zhang X, Liang C, Lu Q, Guo H (2015) Problems with the use of liposome- and retrovirus-mediated gene transfer methods in the primary lymphoid cells of the Oka organs of the greasyback shrimp, Metapenaeus ensis (De Haan, 1844). Crustaceana 88(12–14):1351–1365

    Article  Google Scholar 

  • He F, HoY F, Yu L, Kwang J (2008) WSSV ie1 promoter is more efficient than CMV promoter to express H5 hemagglutinin from influenza virus in baculovirus as a chicken vaccine. BMC Microbiol 8:238

    Article  CAS  Google Scholar 

  • Kafri T, Blömer U, Peterson DA, Gage FH, Verma IM (1997) Sustained expression of genes delivered directly into liver and muscle by lentiviral vectors. Nat Genet 17(3):314–317

    Article  CAS  Google Scholar 

  • Liu ZY, Xiang J, Zhou G, Gong Z (2001) Foreign gene transfer into Chinese shrimps (Penaeus chinensis) with gene gun. Chin Sci Bull 46:766–770

    Article  CAS  Google Scholar 

  • Liu WJ, Chang YS, Wang CH, Kou GH, Lo CF (2005) Microarray and RT-PCR screening for white spot syndrome virus immediate-early genes in cycloheximide-treated shrimp. Virology 334(2):327–341

    Article  CAS  Google Scholar 

  • Lo CF, HO C H, Peng SE, Chen CH, Hsu HC, Chiu YL, Chang HF, Liu KF, Su MS, Wang CH, Kou GH (1996) White spot syndrome baculovirus (WSBV) detected in cultured and captured shrimp, crabs and other arthropods. Dis Aquat Org 27(3):215–225

    Article  Google Scholar 

  • Miyoshi H, Takahashi M, Gage FH, Verma IM (1997) Stable and efficient gene transfer into the retina using an HIV-based lentiviral vector. Proc Natl Acad Sci 94(19):10319–10323

    Article  CAS  Google Scholar 

  • Musthaq SS, Madhan S, Hameed ASS, Kwang J (2009) Localization of VP28 on the baculovirus envelope and its immunogenicity against white spot syndrome virus in Penaeus monodon. Virology 391(2):315–324

    Article  Google Scholar 

  • Naldini L, Blomer U, Gallay P, Ory D, Mulligan R, Gage FH, Verma IM, Trono D (1996) In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 272(5259):263–267

    Article  CAS  Google Scholar 

  • Preston NP, Baule VJ, Leopold R, Henderling J, Atkinson PW, Whyard S (2000) Delivery of DNA to early embryos of the Kuruma prawn, Penaeus japonicas. Aquaculture 181:225–234

    Article  CAS  Google Scholar 

  • Pu L, Wang J, Zhang X, Guo H (2017) Development of pseudotyped retroviral system for effective gene transfer and expression in penaeid shrimp cells. Aquaculture 467:198–210

    Article  CAS  Google Scholar 

  • Puthumana J, Philip R, Singh IB (2016) Transgene expression in Penaeus monodon cells: evaluation of recombinant baculoviral vectors with shrimp specific hybrid promoters. Cytotechnology 68(4):1147–1159

    Article  CAS  Google Scholar 

  • Shike H, Shimizu C, Klimpel KS, Burns JC (2000) Expression of foreign genes in primary cultured cells of the blue shrimp Penaeus stylirostris. Mar Biol 137:605–611

    Article  CAS  Google Scholar 

  • Tang X, Wu J, Sivaraman J, Hew CL (2007) Crystal structures of major envelope proteins VP26 and VP28 from white spot syndrome virus shed light on their evolutionary relationship. J Virol 81(12):6709–6717

    Article  CAS  Google Scholar 

  • Tapay LM, Lu YA, Brock JA, Nadala ECB, Loh PC (1995) Transformation of primary cultures of shrimp (Penaeus stylirostris) lymphoid (Oka) organ with Simian virus-40 (T) antigen. Proc Soc Exp Biol Med 209:73–78

    Article  CAS  Google Scholar 

  • Tsai JM, Wang H-C, Leu J-H, Wang AH-J, Zhuang Y, Walker PJ, Kou G-H, Lo C-F (2006) Identification of the nucleocapsid, tegument, and envelope proteins of the shrimp white spot syndrome virus virion. J Virol 80(6):3021–3029

    Article  CAS  Google Scholar 

  • van Hulten MCW, Westenberg M, Goodall SD, Vlak JM (2000) Identification of two major virion protein genes of white spot syndrome virus of shrimp. Virology 266(2):227–236

    Article  Google Scholar 

  • van Hulten MCW, Witteveldt J, Snippe M, Vlak JM (2001) White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp. Virology 285(2):228–233

    Article  Google Scholar 

  • van Hulten MCW, Reijns M, Vermeesch AMG, Zandbergen F, Vlak JM (2002) Identification of VP19 and VP15 of white spot syndrome virus (WSSV) and glycosylation status of the WSSV major structural proteins. J Gen Virol 83(1):257–265

    Article  Google Scholar 

  • VandenDriessche T, Thorrez L, Naldini L, Follenzi A, Moons L, Berneman Z, Collen D, Chuah MKL (2002) Lentiviral vectors containing the human immunodeficiency virus type-1 central polypurine tract can efficiently transduce nondividing hepatocytes and antigen-presenting cells in vivo. Blood 100(3):813–822

    Article  CAS  Google Scholar 

  • Vigna E, Naldini L (2000) Lentiviral vectors: excellent tools for experimental gene transfer and promising candidates for gene therapy. J Gene Med 2(5):308–316

    Article  CAS  Google Scholar 

  • Yi G, Wang Z, Qi Y, Yao L, Qian J, Hu L (2004) Vp28 of shrimp white spot syndrome virus is involved in the attachment and penetration into shrimp cells. BMB Rep 37(6):726–734

    Article  CAS  Google Scholar 

  • Zufferey R, Nagy D, Mandel RJ, Naldini L, Trono D (1997) Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo. Nat Biotechnol 15:871–875

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank our colleague Dr. Tao Liu for his kind help in the ultra-centrifugation and Dr. Jiankai Wei for his kind help in the confocal observation.

Funding

This work was supported by the National Natural Science Foundation of China (Grant No. 31472274 and 31172391) and Grants 201822018 and 201762003 from the Fundamental Research Funds for Central Universities (China).

Author information

Authors and Affiliations

  1. Ministry of Education Key Laboratory of Marine Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao, 266003, China

    Xuemei Chen, Yueru Chen, Xiaotong Shen, Jianwei Zuo & Huarong Guo

  2. Institute of Evolution and Marine Biodiversity, Ocean University of China, Qingdao, 266003, China

    Huarong Guo

Authors
  1. Xuemei Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yueru Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiaotong Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jianwei Zuo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Huarong Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Huarong Guo.

Ethics declarations

All applicable, international, national, and/or institutional guidelines for the care and use of animals were followed in the present study for shrimps.

Conflict of Interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chen, X., Chen, Y., Shen, X. et al. The Improvement and Application of Lentivirus-Mediated Gene Transfer and Expression System in Penaeid Shrimp Cells. Mar Biotechnol 21, 9–18 (2019). https://doi.org/10.1007/s10126-018-9862-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10126-018-9862-0

Keywords

Search

Navigation