ISSN 0564-3783  

Main page
Information to authors
Editorial board
Mobile version

In Ukrainian

Export citations

Molecular cloning and functional characterization of mouse innate immune sensor RIG

Tangjie Zhang, Saumendra N. Sarkar, Jianzhong Zhu


Innate immunity functions as the first line against infection, which is mediated by series of innate immune receptors. RIG-I like receptors (RLR) recognize the cytosolic nucleic acids mainly from viruses, trigger the type I interferon (IFN) production, and thus play essential roles in the anti-viral immunity. Here we cloned the mouse RIG-I (mRIG-I) cDNA into the pENTR vector using the TOPO cloning technology, and transferred the FLAG tagged mRIG-I gene from pENTR vector to destination pLenti viral vector. The expression of mRIG-I in transfected cells was detected by immunoblotting using anti-FLAG and anti-RIG-I antibodies. The transfected mRIG-I was active capable of inducing downstream IFN stimulated gene (ISG) gene transcription as well as protein expression.

Key words: Gateway cloning; ISG; mouse RIG-I; TOPO cloning

Tsitologiya i Genetika 2019, vol. 53, no. 4, pp. 70-71

  1. College Veterinary Medicine, Yangzhou University, Yangzhou 225009, P. R. China
  2. School of Medicine, University of Pittsburgh, Pittsburgh, USA, 15232

E-mail: slx

Tangjie Zhang, Saumendra N. Sarkar, Jianzhong Zhu Molecular cloning and functional characterization of mouse innate immune sensor RIG, Tsitol Genet., 2019, vol. 53, no. 4, pp. 70-71.

In "Cytology and Genetics":
Tangjie Zhang, Saumendra N. Sarkar, Jianzhong Zhu Molecular Cloning and Functional Characterization of Mouse Innate Immune Sensor RIG-I, Cytol Genet., 2019, vol. 53, no. 4, pp. 325329
DOI: 10.3103/S0095452719040121


1. ONeill, L.A., Immunology. After the Toll rush, Science, 2004, vol. 303, pp. 14811482.

2. Medzhitov, R., Approaching the asymptote: 20 years later, Immunity, 2009, vol. 30, pp. 766775.

3. Schlee, M., Master sensors of pathogenic RNARIG-I like receptors, Immunobiology, 2013, vol. 218, pp. 13221335.

4. Hornung, V., Ellegast, J., Kim, S., Brzozka, K., Jung, A., Kato, H., Poeck, H., Akira, S., Conzelmann, K.K., Schlee, M., Endres, S., and Hartmann, G., 5'-triphosphate RNA is the ligand for RIG-I, Science, 2006, vol. 314, pp. 994997.

5. Bowie, A.G. and Fitzgerald, K.A., RIG-I: tri-ing to discriminate between self and non-self RNA, Trends Immunol., 2007, vol. 28, pp. 147150.

6. Zhu, J., Ghosh, A., and Sarkar, S.N., OASLa new player in controlling antiviral innate immunity, Curr. Opin. Virol., 2015, vol. 12, pp. 1519.

7. Yoneyama, M., Kikuchi, M., Natsukawa, T., Shinobu, N., Imaizumi, T., Miyagishi, M., Taira, K., Akira, S., and Fujita, T., The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses, Nat. Immunol., 2004, vol. 5, pp. 730737.

8. Sumpter, R., Jr., Loo, Y.M., Foy, E., Li, K., Yoneyama, M., Fujita, T., Lemon, S.M., and Gale, M., Jr., Regulating intracellular antiviral defense and permissiveness to hepatitis C virus RNA replication through a cellular RNA helicase, RIG-I, J. Virol., 2005, vol. 79, pp. 26892699.

9. Kato, H., Takeuchi, O., Sato, S., Yoneyama, M., Yamamoto, M., Matsui, K., Uematsu, S., Jung, A., Kawai, T., Ishii, K.J., Yamaguchi, O., Otsu, K., Tsujimura, T., Koh, C.S., Reis e Sousa, C., Matsuura, Y., Fujita, T., and Akira, S., Differential roles of MDA5 and RIG-I helicases in the recognition of RNA viruses, Nature, 2006, vol. 441, pp. 101105.

10. Kell, A.M. and Gale, M., Jr., RIG-I in RNA virus recognition, Virology, 2015, vol. 479-80, pp. 110121.

11. Wu, B. and Hur, S., How RIG-I like receptors activate MAVS, Curr. Opin. Virol., 2015, vol. 12, pp. 9198.

12. Zhu, J., Zhang, Y., Ghosh, A., Cuevas, R.A., Forero, A., Dhar, J., Ibsen, M.S., Schmid-Burgk, J.L., Schmidt, T., Ganapathiraju, M.K., Fujita, T., Hartmann, R., Barik, S., Hornung, V., Coyne, C.B., and Sarkar, S.N., Antiviral activity of human OASL protein is mediated by enhancing signaling of the RIG-I RNA sensor, Immunity, 2014, vol. 40, pp. 936948.

13. Campeau, E., Ruhl, V.E., Rodier, F., Smith, C.L., Rahmberg, B.L., Fuss, J.O., Campisi, J., Yaswen, P., Cooper, P.K., and Kaufman, P.D., A versatile viral system for expression and depletion of proteins in mammalian cells, PLoS One, 2009, vol. 4, no. 8. e6529.

14. Esposito, D., Garvey, L.A., and Chakiath, C.S., Gateway cloning for protein expression, Methods Mol. Biol., 2009, vol. 498, pp. 3154.

15. Meerbrey, K.L., Hu, G., Kessler, J.D., Roarty, K., Li, M.Z., Fang, J.E., Herschkowitz, J.I., Burrows, A.E., Ciccia, A., Sun, T., Schmitt, E.M., Bernardi, R.J., Fu, X., Bland, C.S., Cooper, T.A., Schiff, R., Rosen, J.M., Westbrook, T.F., and Elledge, S.J., The pINDUCER lentiviral toolkit for inducible RNA interference in vitro and in vivo, Proc. Natl. Acad. Sci. U. S. A., 2011, vol. 108, pp. 36653670.

Copyright© ICBGE 2002-2022 Coded & Designed by Volodymyr Duplij Modified 17.05.22