Share This Article:

Pronuclear microinjection is not suitable for RNA polymerase III promoter driven constitutive RNAi transgenesis in mice for XY male-to-female sex reversal by Sry gene knockdown

Abstract Full-Text HTML Download Download as PDF (Size:727KB) PP. 62-69
DOI: 10.4236/ojgen.2012.21008    4,762 Downloads   8,151 Views  
Author(s)    Leave a comment

ABSTRACT

Silencing of gene expression by RNA interference (RNAi) has become a widely used tool. For the study of mammalian gene function expression vectors for short hairpin RNA (shRNA) were developed. However the standard methods of shRNA transgenic (Tg) mice production have not been established. Sry (sex-determining region on the Y chromosome) is a mammalian sex-determining gene on the Y chromosome. In mice, the transient expression of Sry in supporting cell precursor cells between 10.5 and 12.5 days post-coitus (dpc) triggers the differentiation of Sertoli cells from granulosa cells. Then high efficiency of Sry gene silencing in Tg mice should induce XY male-to-female sex reversal. An shRNA Tg mouse targeting Sry gene was attempted to be generated by pronuclear microinjection. A low rate (Tg pups/all pups born after microinjection = 2/154 to 7/178) of Tg pups was observed. These Tg mice showed no XY male-to-female sex reversal. The results suggest that exogenous expression of small RNA might exert a negative effect on embryonic development and another approach should be needed for RNAi transgenesis in mice.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

Ito, M. (2012) Pronuclear microinjection is not suitable for RNA polymerase III promoter driven constitutive RNAi transgenesis in mice for XY male-to-female sex reversal by Sry gene knockdown. Open Journal of Genetics, 2, 62-69. doi: 10.4236/ojgen.2012.21008.

References

[1] Ito, M., Zhao, N., Zeng, Z, Chang, C.C. and Zu, Y. (2010) Synergistic growth inhibition of anaplastic large cell lymphoma cells by combining cellular ALK gene silencing and a low dose of the kinase inhibitor U0126. Cancer Gene Therapy, 17, 633-644. doi:10.1038/cgt.2010.20
[2] Ito, M., Kawano, K., Miyagishi, M. and Taira, K. (2005) Genome-wide application of RNAi to the discovery of potential drug targets. FEBS Letters, 579, 5988-5995. doi:10.1016/j.febslet.2005.08.015
[3] Koopman, P., Gubbay, J., Vivian, N., Goodfellow, P. and Lovell-Badge, R. (1991) Male development of chromosomally female mice transgenic for Sry. Nature, 351, 117-121. doi:10.1038/351117a0
[4] Bullejos, M. and Koopman, P. (2001) Spatially dynamic expression of Sry in mouse genital ridges. Developmental Dynamics, 221, 201-205. doi:10.1002/dvdy.1134
[5] Albrecht, K.H. and Eicher, E.M. (2001) Evidence that Sry is expressed in pre-Sertoli cells and Sertoli and granulosa cells have a common precursor. Developmental Biology, 240, 92-107. doi:10.1006/dbio.2001.0438
[6] Ito, M., Yokouchi, K., Yoshida, K., Kano, K., Naito, K., Miyazaki, J. and Tojo, H. (2006) Investigation of the fate of Sry-expressing cells using an in vivo Cre/loxP system. Development Growth and Differentiation, 48, 41-47. doi:10.1046/j.1440-169X.2002.00668.x
[7] Ito, M., Miyagishi, M., Murata, C., Kawasaki, H., Baba, T., Tachi, C. and Taira, K. (2006) Down-regulation of endogenous Wt1 expression by Sry transgene in the murine embryonic mesonephros-derived M15 cell line. Journal of Reproduction and Development, 52, 415-427. doi:10.1262/jrd.17094
[8] Hogan, B., Constantini, F. and Lacy, E. (1994) Manipulating the mouse embryo. 2nd Edition, Cold Spring Harbor Laboratory Press, New York.
[9] Yoshida, K., Ito, M., Yokouchi, K., Kano, K., Naito, K. and Tojo H. (2010) Ubiquitous expression of Sry induces embryonic lethality related to suppression of Tie2/Tek expression. Advances in Bioscience and Biotechnology, 1, 444-452. doi:10.4236/abb.2010.15058
[10] Kato, M., Yamanouchi, K., Ikawa, M., Okabe, M., Naito, K. and Tojo, H. (1999) Efficient selection of transgenic mouse embryos using EGFP as a marker gene. Molecular Reproduction and Development, 54, 43-48. doi:10.1002/(SICI)1098-2795(199909)54:1<43::AID-MRD6>3.0.CO;2-N
[11] Seo, B.B., Kim, C.H., Yamanouchi, K., Takahashi, M., Sawasaki, T., Tachi, C. and Tojo, H. (2000) Coinjection of restriction enzyme with foreign DNA into the pronucleus for elevating production efficiencies of transgenic animals. Animal Reproduction Science, 63, 113-122. doi:10.1016/S0378-4320(00)00148-2
[12] Kubo, J., Yamanouchi, K., Naito, K. and Tojo, H. (2002) Expression of the gene of interest fused to the EGFP-expressing gene in transgenic mice derived from selected transgenic embryos. Journal of Experimental Zoology, 293, 712-718. doi:10.1002/jez.10163
[13] Ito, M., Yamanouchi, K., Naito, K. Calos, M.P. and Tojo, H. (2011) Site-specific integration of transgene targeting an endogenous lox-like site in early mouse embryos. Journal of Applied Genetics, 52, 89-94. doi:10.1007/s13353-010-0011-3
[14] Svingen, T., Wilhelm, D., Combes, A.N., Hosking, B., Harley, V.R., Sinclair, A.H. and Koopman, P. (2009) Ex vivo magnetofection: A novel strategy for the study of gene function in mouse organogenesis. Developmental Dynamics, 238, 956-964. doi:10.1002/dvdy.21919
[15] Wu, N., Lin, X.K., Liao, B., Du, W.H., Han, F.T. and Zhao, J.H. (2008) Effect of Sry silencing by siRNA on the expression of sex determining genes in mouse embryos. Yi Chuan, 30, 195-202. doi:10.3724/SP.J.1005.2008.00195
[16] Hasuwa, H., Kaseda, K., Einarsdottir, T. and Okabe, M. (2002) Small interfering RNA and gene silencing in transgenic mice and rats. FEBS Letters, 532, 227-230. doi:10.1016/S0014-5793(02)03680-3
[17] Carmell, M.A., Zhang, L., Conklin, D.S., Hannon, G.J. and Rosenquist, T.A. (2003) Germline transmission of RNAi in mice. Nature Structural Biology, 10, 91-92. doi:10.1038/nsb896
[18] Peng, S., York, J.P. and Zhang, P. (2006) A transgenic approach for RNA interference-based genetic screening in mice. Proceedings of the National Academy of Sciences of USA, 103, 2252-2256. doi:10.1073/pnas.0511034103
[19] Shinagawa, T. and Ishii, S. (2003) Generation of Skiknockdown mice by expressing a long double-strand RNA from an RNA poly-merase II promoter. Genes & Development, 17, 1340-1345. doi:10.1101/gad.1073003
[20] Stein, P., Svoboda, P. and Schultz, R.M. (2003) Transgenic RNAi in mouse oocytes: A simple and fast approach to study gene function. Developmental Biology, 256, 187-193. doi:10.1016/S0012-1606(02)00122-7
[21] Yu, J., Deng, M., Medvedev, S., Yang, J., Hecht, N.B. and Schultz, R.M. (2004) Transgenic RNAi-mediated reduction of MSY2 in mouse oocytes results in reduced fertility. De-velopmental Biology, 268, 195-206. doi:10.1016/j.ydbio.2003.12.020
[22] Stein, P., Zeng, F., Pan, H. and Schultz, R.M. (2005) Absence of non-specific effects of RNA interference triggered by long double-stranded RNA in mouse oocytes. Developmental Biology, 286, 464-471. doi:10.1016/j.ydbio.2005.08.015
[23] Sarnova, L., Malik, R., Sedlacek, R. and Svoboda, P. (2010) Shortcomings of short hairpin RNA-based transgenic RNA interference in mouse oocytes. Journal of Negative Results in Biomedicine, 9, 8. doi:10.1186/1477-5751-9-8
[24] Coumoul, X., Shukla, V., Li, C., Wang, R.H. and Deng, C. (2004) Conditional knockdown of Fgfr2 in mice using Cre-LoxP induced RNA interference. Nucleic Acids Research, 33, e102. doi:10.1093/nar/gni100
[25] Dickins, R.A., McJunkin, K., Hernando, E., Premsrirut, P.K., Krizhanovsky, V., Burgess, D.J., Kim, S.Y., Cordon-Cardo, C., Zender, L., Hannon, G.J. and Lowe, S.W. (2007) Tissue-specific and reversible RNA interference in transgenic mice. Nature Genetics, 39, 914-921. doi:10.1038/ng2045
[26] Pfeifer, A. (2004) Lentiviral transgenesis. Transgenic Research, 13, 513-522. doi:10.1007/s11248-004-2735-5
[27] Tiscornia, G., Singer, O., Ikawa, M. and Verma, I.M. (2003) A general method for gene knockdown in mice by using lentiviral vectors expressing small interfering RNA. Proceedings of the National Academy of Sciences of USA, 100, 1844-1848. doi:10.1073/pnas.0437912100
[28] Rubinson, D.A., Dillon, C.P., Kwiatkowski, A.V., Sievers, C., Yang, L., Kopinja, J., Rooney, D.L., Zhang, M., Ihrig, M.M., McManus, M.T., Gertler, F.B., Scott, M.L. and Van Parijs, L. (2003) A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference. Nature Genetics, 33, 401-406. doi:10.1038/ng1117
[29] Kirilov, M., Chai, M., van der Hoeven, F., Kloz, U., Schmid, W. and Schütz, G. (2007) Germ line transmission and expression of an RNAi cassette in mice generated by a lentiviral vector system. Transgenic Research, 16, 783- 793. doi:10.1007/s11248-007-9119-6
[30] Kunath, T., Gish, G., Lickert, H., Jones, N., Pawson, T. and Rossant, J. (2003) Transgenic RNA interference in ES cell-derived embryos recapitulates a genetic null phenotype. Nature Biotechnology, 21, 559-561. doi:10.1038/nbt813
[31] Lickert, H., Cox, B., Wehrle, C., Taketo, M.M., Kemler, R. and Rossant, J. (2005) Dissecting Wnt/betacatenin signaling during gastrulation using RNA interference in mouse embryos. Development, 132, 2599-2609. doi:10.1242/dev.01842
[32] Ventura, A., Meissner, A., Dillon, C.P., McManus, M., Sharp, P.A., Van Parijs, L., Jaenisch, R. and Jacks, T. (2004) Crelox-regulated conditional RNA interference from transgenes. Proceedings of the National Academy of Sciences of USA, 101, 10380-10385. doi:10.1073/pnas.0403954101
[33] Stern, P., Astrof, S., Erkeland, S.J., Schustak, J., Sharp, P.A. and Hynes, R.O. (2008) A system for Creregulated RNA interference in vivo. Proceedings of the National Academy of Sciences of USA, 105, 13895-13900. doi:10.1073/pnas.0806907105
[34] Seibler, J., Kleinridders, A., Küter-Luks, B., Niehaves, S., Brüning, J.C. and Schwenk, F. (2007) Reversible gene knockdown in mice using a tight, inducible shRNA expression system. Nucleic Acids Research, 35, e54. doi:10.1093/nar/gkm122
[35] Vidigal, J.A., Morkel, M., Wittler, L., Brouwer-Lehmitz, A., Grote, P., Macura, K. and Herrmann, B.G. (2010) An inducible RNA interference system for the functional dissection of mouse embryogenesis. Nucleic Acids Research, 38, e122. doi:10.1093/nar/gkq199
[36] Premsrirut, P.K., Dow, L.E., Kim, S.Y., Camiolo, M., Malone, C.D., Miething, C., Scuoppo, C., Zuber, J., Dickins, R.A., Kogan, S.C., Shroyer, K.R., Sordella, R., Hannon, G.J. and Lowe, S.W. (2011) A rapid and scalable system for studying gene function in mice using conditional RNA interference. Cell, 145, 145-158. doi:10.1016/j.cell.2011.03.012
[37] McJunkin, K., Ma-zurek, A., Premsrirut, P.K., Zuber, J., Dow, L.E., Simon, J., Stillman, B. and Lowe, S.W (2011) Reversible suppression of an essential gene in adult mice using transgenic RNA interference. Proceedings of the National Academy of Sciences of USA, 108, 7113-7118. doi:10.1073/pnas.1104097108
[38] Svoboda, P. (2004) Long dsRNA and silent genes strike back: RNAi in mouse oocytes and early embryos. Cytogenetic and Genome Research, 105, 422-434. doi:10.1159/000078215
[39] Ito, M., Yokouchi, K., Naito, K., Endo, H., Hakamata, Y., Miyazaki, J. and Tojo, H. (2002) In vitro Cre/loxP system in cells from developing gonads: Investigation of the Sry promoter. Development Growth and Differentiation, 44, 549-557. doi:10.1046/j.1440-169X.2002.00668.x
[40] Ito, M., Yokouchi, K., Naito, K., Endo, H., Hakamata, Y., Miya-zaki, J. and Tojo, H. (2005) Detection of elements responsible for stage- and tissue-specific expression of mouse Sry using an in vitro Cre/loxP system. Biochemical and Biophysical Research Communications, 337, 264- 270. doi:10.1016/j.bbrc.2005.09.047
[41] Beard, C., Hochedlinger, K., Plath, K., Wutz, A. and Jaenisch, R. (2006) Efficient method to generate singlecopy transgenic mice by site-specific integration in embryonic stem cells. Genesis, 44, 23-28. doi:10.1002/gene.20180
[42] Kleinhammer, A., Wurst, W. and Kühn, R. (2011) Constitutive and conditional RNAi transgenesis in mice. Methods, 53, 430-436. doi:10.1016/j.ymeth.2010.12.015
[43] Kleinhammer, A., Deussing, J., Wurst, W. and Kühn, R. (2011) Conditional RNAi in mice. Methods, 53, 142-150. doi:10.1016/j.ymeth.2010.08.003

  
comments powered by Disqus

Copyright © 2019 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.