Design of hammerhead ribozymes that cleave murine sry mrna in vitro and in vivo

Kaori Nakamura, Chisato Murata, Masanori Ito, Tokuko Iwamori, Seiichiro Nishimura, Shin Hisamatsu, Shigenori Sonoki, Aya Nakayama, Eigo Suyama, Hiroaki Kawasaki, Kazunari Taira, Koichiro Nishino, Chikashi Tachi

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Abstract

As the first step in investigating the possiblity of applying ribozyme technology to artificial control of the sex ratios at birth in farm animals, where the demand for females exceeds that for males, we designed a hammerhead ribozyme (HHRz) and 2 tRNA val-hammerhead ribozyme complexes (tRNARzS and tRNARz4), and examined their effects upon murine Sry mRNA in vitro and in cells. We demonstrated that HHRz and tRNARz3 could effectively cleave the target Sry mRNA in vitro. For the purpose of experiments in vivo, HHRz was cloned into the highly efficient pUC-CAGGS mammalian expression vector (pCAG/HHRz), and the tRNA ribozyme complexes were cloned into the pol III promoter-driven pPUR-KE vector (pPUR/tRNARz3 and pPUR/tRNARz4); the ribozyme vectors were co-transfected with the target vector (pCAG/Sry). A suppressive action (up to approx. 60%) was confirmed for pCAG/HHRz and pPUR/tRNARz3 upon the transiently expressed exogenously introduced Sry in M15 cultured cells.

Original languageEnglish
Pages (from-to)73-80
Number of pages8
JournalJournal of Reproduction and Development
Volume52
Issue number1
DOIs
Publication statusPublished - Feb 2006

All Science Journal Classification (ASJC) codes

  • Animal Science and Zoology

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    Nakamura, K., Murata, C., Ito, M., Iwamori, T., Nishimura, S., Hisamatsu, S., Sonoki, S., Nakayama, A., Suyama, E., Kawasaki, H., Taira, K., Nishino, K., & Tachi, C. (2006). Design of hammerhead ribozymes that cleave murine sry mrna in vitro and in vivo. Journal of Reproduction and Development, 52(1), 73-80. https://doi.org/10.1262/jrd.17060