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Using a nano-flare probe to detect RNA in live donor cells prior to somatic cell nuclear transfer

文献类型： 外文期刊

作者： Fu, Bo 1 ; Ren, Liang 3 ; Liu, Di 2 ; Ma, Jian-zhang 1 ; An, Tie-zhu 1 ; Yang, Xiu-qin 3 ; Ma, Hong 2 ; Guo, Zhen-hua 2 ; Zh 1 ;

作者机构： 1.Northeast Forestry Univ, Heilongjiang Acamedy Agr Sci Postdoctoral Program, Postdoctoral Programme, Harbin, Peoples R China

2.HeiLongJiang Acad Agr Sci, Inst Anim Husb Res, Harbin 150086, Peoples R China

3.Northeast Agr Univ, Coll Anim Sci, Harbin 150030, Peoples R China

4.Heilongjiang Univ Chinese Med, Modern Educ Technol & Informat Ctr

关键词： development;nanoparticle;RNA detection;somatic cell nuclear transfer

期刊名称：CELL BIOLOGY INTERNATIONAL （ 影响因子：3.612； 五年影响因子：3.055 ）

ISSN：

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收录情况： SCI

摘要： Many transgenes are silenced in mammalian cells (donor cells used for somatic cell nuclear transfer [SCNT]). Silencing correlated with a repressed chromatin structure or suppressed promoter, and it impeded the production of transgenic animals. Gene transcription studies in live cells are challenging because of the drawbacks of reverse-transcription polymerase chain reaction and fluorescence in situ hybridization. Nano-flare probes provide an effective approach to detect RNA in living cells. We used 18S RNA, a housekeeping gene, as a reference gene. This study aimed to establish a platform to detect RNA in single living donor cells using a Nano-flare probe prior to SCNT and to verify the safety and validity of the Nano-flare probe in order to provide a technical foundation for rescuing silenced transgenes in transgenic cloned embryos. We investigated cytotoxic effect of the 18S RNA-Nano-flare probe on porcine fetal fibroblasts, characterized the distribution of the 18S RNA-Nano-flare probe in living cells and investigated the effect of the 18S RNA-Nano-flare probe on the development of cloned embryos after SCNT. The cytotoxic effect of the 18S RNA-Nano-flare probe on porcine fetal fibroblasts was dose-dependent, and 18S RNA was detected using the 18S RNA-Nano-flare probe. In addition, treating donor cells with 500pM 18S RNA-Nano-flare probe did not have adverse effects on the development of SCNT embryos at the pre-implantation stage. In conclusion, we established a preliminary platform to detect RNA in live donor cells using a Nano-flare probe prior to SCNT.

  • 相关文献

[1]Effect of Pyruvate and Lactic Acid on the Development of Porcine Embryos. Fu Bo,Llu Di,Ma Hong,Li Zhong-qiu,Wang Liang,Bai Jing. 2012

[2]Effect of Cycloheximide on the Development of Parthenogenetic Activation and Cloned Porcine Embryos. Ma, Hong,Fu, Bo,Li, Zhongqiu,Liu, Di,Zhao, Jinfeng,Fang, Qingchang,Ren, Liang. 2011

[3]Development of porcine tetraploid somatic cell nuclear transfer embryos is influenced by oocyte nuclei. Fu, Bo,Liu, Di,Ma, Hong,Guo, Zhen-Hua,Wang, Liang,Li, Zhong-Qiu,Peng, Fu-Gang,Liu, Di,Bai, Jing.

[4]Effect of trichostatin A on transfected donor cells and subsequent development of porcine cloned embryos. Liu Di,Ma Hong,Wang Liang,Guo Zhen-hua,Li Zhong-qiu,Fu Bo,Liu Di,Fang Qing-chang,Ren Liang.

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