Evaluation of availability of various internal ribosome entry sites (IRESs) for bicistronic expression in different fish cell lines
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The internal ribosome entry site (IRES) elements from various RNA viruses are widely used to express proteins in bicistronic or multicistronic ways in mammalian cells. However, research on the application of IRESs in fish cells has been poorly conducted. In this study, to evaluate the availability of various viral IRESs in fish cells based on a bicistronic vector system, the translation activity of IRESs from viruses of invertebrates (aphid lethal paralysis virus ALPV; cricket paralysis virus, CrPV; and Plautia stali intestine virus, PSIV), of mammals (encephalomyocarditis virus, EMCV), and of fish (infectious pancreatic necrosis virus, IPNV; marine birnavirus, MABV; and snakehead retrovirus, SnRV) was analyzed in various fish cell lines originated from salmonid (chinook salmon embryonic cells, CHSE-214), cyprinid (Epithelioma papulosum cyprinid, EPC), and flatfish (Hirame natural embryonic cells, HINAE). Translation mediated by EMCV IRES was shown in all cell lines, but the activity was weak in CHSE-214 and HINAE cells while high activity was shown in EPC cells, suggesting that although EMCV IRES can be broadly used in various fish cells, the translational activity can be varied according to different cell lines. PSIV IRES showed moderate activity in EPC cells and low activity in CHSE-214 and HINAE cells. CrPV IRES showed weak activity in EPC and HINAE cells and no activity in CHSE-214 cells. In the case of ALPV IRES, green fluorescence was observed only in EPC cells but the activity was very weak. These results suggest that dicistroviral IRESs analyzed in this study would not be the best option for protein expression in various fish cells. The translation activity of SnRV IRES in fish cells was first verified in EPC cells, though the activity was very weak. The 5’UTR of MABV segment A showed high translation activities in all examined fish cell lines, suggesting that IRES of MABV can be used to develop multicistronic expression systems or cap-independent RNA-based translation systems in a wide range of fish cells.
Abstract
The internal ribosome entry site (IRES) elements from various RNA viruses are widely used to express proteins in bicistronic or multicistronic ways in mammalian cells. However, research on the application of IRESs in fish cells has been poorly conducted. In this study, to evaluate the availability of various viral IRESs in fish cells based on a bicistronic vector system, the translation activity of IRESs from viruses of invertebrates (aphid lethal paralysis virus ALPV; cricket paralysis virus, CrPV; and Plautia stali intestine virus, PSIV), of mammals (encephalomyocarditis virus, EMCV), and of fish (infectious pancreatic necrosis virus, IPNV; marine birnavirus, MABV; and snakehead retrovirus, SnRV) was analyzed in various fish cell lines originated from salmonid (chinook salmon embryonic cells, CHSE-214), cyprinid (Epithelioma papulosum cyprinid, EPC), and flatfish (Hirame natural embryonic cells, HINAE). Translation mediated by EMCV IRES was shown in all cell lines, but the activity was weak in CHSE-214 and HINAE cells while high activity was shown in EPC cells, suggesting that although EMCV IRES can be broadly used in various fish cells, the translational activity can be varied according to different cell lines. PSIV IRES showed moderate activity in EPC cells and low activity in CHSE-214 and HINAE cells. CrPV IRES showed weak activity in EPC and HINAE cells and no activity in CHSE-214 cells. In the case of ALPV IRES, green fluorescence was observed only in EPC cells but the activity was very weak. These results suggest that dicistroviral IRESs analyzed in this study would not be the best option for protein expression in various fish cells. The translation activity of SnRV IRES in fish cells was first verified in EPC cells, though the activity was very weak. The 5’UTR of MABV segment A showed high translation activities in all examined fish cell lines, suggesting that IRES of MABV can be used to develop multicistronic expression systems or cap-independent RNA-based translation systems in a wide range of fish cells.
참고문헌 (30)
- Attal, J., "The opti-mal use of IRES(internal ribosome entry site)in expression vectors", Genet Anal, 15, pp.161~165 (1999)
- Bochkov, Y. A., "Translational efficiency of EMCV IRES in bicistronic vectors is dependent upon IRES sequence and gene location", BioTechniques, 41 (2006)
- Bonning, B. C., "Dicistroviruses", Annu Rev Entomol, 55, pp.129~150 (2010)
- Borman, A. M., "Picornavirus internal ribosome entry segments : comparison of translation efficiency and the require-ments for optimal internal initiation of translation in vitro", Nucleic Acids Res, 23, pp.3656~3663 (1995)
- de Breyne, S., "Focus on Translation Initiation of the HIV-1 mRNAs", Int J Mol Sci, 20, pp.101~ (2018)
- Delmas, B., "ICTV virus taxonomy profile: Birnaviridae", J Gen Virol, 100, pp.5~6 (2019)
- Derrington, E. A., "Dicistronic MLV-retroviral vectors transduce neural precursors in vivo and co-express two genes in their differentiated neuronal progeny", Retrovirology, 2, pp.60~ (2005)
- Dobos P, "In vitro guanylylation of infectious pancreatic necrosis virus polypeptide VP1", Virology, 193, pp.403~413 (1993)
- Fahrenkrug, S. C., "Dicistronic Gene Expression in Developing Zebrafish", Mar Biotechnol(NY), 1, pp.552~561 (1999)
- Fukamachi, S., "Effects of constitutive expression of somatolactin al- pha on skin pigmentation in medaka", Gene, 442, pp.81~87 (2009)
- Gamil, A. A., "A piscine birnavirus induces inhibition of protein synthesis in CHSE-214 cells primarily through the induction of eIF2α phosphorylation", Viruses, 7, pp.1987~2005 (2015)
- Godet, A. C., "IRES Trans-Acting Factors, Key Actors of the Stress Response", Int J Mol Sci, 20, pp.924~ (2019)
- Hart, D., "Complete nucleotide sequence and transcriptional analysis of snakehead fish retrovirus", J Virol, 70, pp.3606~3616 (1996)
- Hertz, M. I., "In vivo functional analysis of the Dicistroviridae intergenic region internal ribosome entry sites", Nucleic Acids Res, 39, pp.7276~7288 (2011)
- Jaafar, Z. A., "Viral RNA structure-based strategies to manipulate translation", Nat Rev Micro-biol, 17, pp.110~123 (2019)
- Jang, S. K., "A seg-ment of the 5' nontranslated region of encephalo-myocarditis virus RNA directs internal entry of ribo-somes during in vitro translation", J Virol, 62, pp.2636~2643 (1988)
- Johnson, A. G., "Dynamics of IRES-mediated translation", Philos Trans R Soc Lond B Biol Sci, 372 (2017)
- Kim So Yeon, "Marine birnavirus (MABV)’s 5´ terminal region of segment A acts as internal ribosome entry site (IRES)", 한국어병학회지, 34(1), pp.17~22 (2021), 한국어병학회
- Komar, A. A., "Cellular IRES-medi- ated translation: the war of ITAFs in pathophysiological states", Cell cycle, 10, pp.229~240 (2011)
- Licursi, M., "Promotion of Viral IRES-Mediated Translation Initiation under Mild Hypothermia", PLoS One, 10, pp.e0126174~ (2015)
- Liu, L., "Gen-eration and functional evaluation of a DNA vaccine co-expressing Cyprinid herpesvirus-3 envelope protein and carp interleukin-1 beta", Fish Shellfish Im-munol, 80, pp.223~231 (2018)
- Lozano, G., "Structural insights into viral IRES-dependent translation mechanisms", Curr Opin Virol, 12, pp.113~120 (2015)
- Lu, J., "In vitro and in vivo identification of structural and sequence elements in the 5' untranslated region of Ectropis obliqua picorna-like virus required for internal initiation", J Gen Virol, 87, pp.3667~3677 (2006)
- Martínez-Salas, E., "Picornavirus IRES elements : RNA structure and host protein interactions", Virus Res, 206, pp.62~73 (2015)
- Pelletier, J., "Internal initiation of translation of eukaryotic mRNA directed by a se- quence derived from poliovirus RNA", Nature, 334, pp.320~325 (1988)
- Rivas-Aravena, A., "Study of RNA-A Initiation Translation of The Infectious Pancreatic Necrosis Virus", Virus Res, 240, pp.121~129 (2017)
- Sonenberg, N., "Regulation of translation initiation in eukaryotes: mechanisms and biological targets", Cell, 136, pp.731~745 (2009)
- Spriggs, K. A., "Re-programming of translation following cell stress allows IRES-mediated translation to predominate", Biol Cell, 100, pp.27~38 (2008)
- Walsh, D., "Viral subversion of the host protein synthesis machinery", Nat Rev Microbiol, 9, pp.860~875 (2011)
- Xu, H., "Generation and functional evaluation of a DNA vac-cine co-expressing Vibrio anguillarum VAA protein and flounder interleukin-2", Fish Shellfish Immunol, 93, pp.1018~1027 (2019)