Kwon Sujin, Jeong Sangkyun, Jeong Young Sun, Park Jung Sun, Cui Xiang-Shun, Kim Nam-Hyung, Kang Yong-Kook
1 Epigenetics Research Center , KRIBB, Daejeon, 305-806, Korea.
Cell Reprogram. 2015 Feb;17(1):49-58. doi: 10.1089/cell.2014.0057. Epub 2014 Dec 30.
Researchers have exerted sustained efforts to improve the viability of somatic cell nuclear transfer (SCNT) embryos, testing their experimental designs and probing the resultant embryos. However, the lack of a reliable method to estimate the efficacy of these experimental attempts is a chief hindrance to tackling the low-viability problem in SCNT. Here, we introduce a procedure that assesses the degree of difference in gene expression profiles (GEPs) of blastocysts from each other as a representative control of good quality. We first adapted a multiplex reverse transcription-polymerase chain reaction strategy to obtain GEPs for 15 reprogramming-related genes from single mouse blastocysts. GEPs of individual blastocysts displayed a broad range of variations, the extent of which was calculated using a weighted root mean square deviation (wRMSD). wRMSD-based quantitation of GEP difference (qGEP) found that GEP difference between in vivo-derived blastocysts (in vivo) and SCNT blastocysts was greater than the difference between in vivo blastocysts and in vitro-produced (IVP) blastocysts, demonstrating that the SCNT group was more distantly related to the in vivo group than the IVP group. Our qGEP approach for grading individual blastocysts would be useful for selecting a better protocol to derive embryos of better quality prior to field applications.
研究人员一直在持续努力提高体细胞核移植（SCNT）胚胎的存活率，对他们的实验设计进行测试并探究所得胚胎。然而，缺乏一种可靠的方法来评估这些实验尝试的效果是解决SCNT低存活率问题的主要障碍。在此，我们引入一种程序，该程序将评估来自彼此的囊胚的基因表达谱（GEP）差异程度作为优质代表性对照。我们首先采用多重逆转录 - 聚合酶链反应策略从单个小鼠囊胚中获取15个重编程相关基因的GEP。单个囊胚的GEP表现出广泛的变化，其程度使用加权均方根偏差（wRMSD）进行计算。基于wRMSD的GEP差异定量（qGEP）发现，体内来源囊胚（体内）与SCNT囊胚之间的GEP差异大于体内囊胚与体外生产（IVP）囊胚之间的差异，这表明SCNT组与体内组的关系比IVP组更远。我们用于对单个囊胚进行分级的qGEP方法将有助于在实际应用之前选择更好的方案来获得质量更好的胚胎。
