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

第一作者： Fu, Bo

作者： Fu, Bo;Ma, Jian-zhang;An, Tie-zhu;Fu, Bo;Liu, Di;Ma, Hong;Guo, Zhen-hua;Ren, Liang;Liu, Di;Yang, Xiu-qin;Zhu, Meng;Bai, Jing

作者机构：

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 ）

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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.

分类号： Q2

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