1] Imagene, R&D Department, Université de Bordeaux 2, ENSTBB, Bordeaux, France [2] Imagene, Production Platform, Rue Henri Desbruères, Evry, France.
Imagene, Production Platform, Rue Henri Desbruères, Evry, France.
Eur J Hum Genet. 2014 Mar;22(3):379-85. doi: 10.1038/ejhg.2013.145. Epub 2013 Jul 17.
RNA is a tool used in many fields, from molecular and cellular biology to medicine and nanotechnology. For most of these uses, the integrity of RNA is required and must be maintained during storage. Even though freezing is currently the storage method of choice, the increasing number of samples to be stored and the costly use of a cold chain have highlighted the need for room temperature preservation methods. Here, we report a new room temperature technology that consists in drying RNA samples in the presence of a stabilizer in stainless steel minicapsules. These air- and water-tight capsules isolate RNA from the atmosphere and maintain an anhydrous and anoxic environment. Through the evaluation of RNA integrity over time at room temperature or 90 °C, we identified atmospheric humidity as a major deleterious factor. The degradation rate dependence in temperature fitted an Arrhenius model, with an activation energy of 28.5 kcal/mol and an extrapolated room temperature degradation rate of 3.2 10(-13)/nt/s (95% confidence interval: 2.3-4.2/nt/s). In these conditions, it is expected that an RNA molecule will be subjected to 0.7-1.3 cut every 1000 nucleotides per century. In addition, we showed that stored RNA is compatible for further analyses, such as reverse transcription-quantitative PCR. No significant change in the Cq values was observed over a simulated period of several decades. At last, our data are consistent with a sequence-independent degradation rate of RNA in the solid state.
RNA 是一种在许多领域都有应用的工具,从分子和细胞生物学到医学和纳米技术。对于大多数这些用途,都需要 RNA 的完整性,并在储存过程中保持其完整性。尽管冷冻目前是首选的储存方法,但需要储存的样本数量不断增加,以及冷链的昂贵使用,突出了需要开发室温保存方法。在这里,我们报告了一种新的室温技术,该技术包括在不锈钢微型胶囊中稳定剂存在的情况下干燥 RNA 样品。这些空气和水密胶囊将 RNA 与大气隔离开来,并保持无水和缺氧环境。通过在室温或 90°C 下随时间评估 RNA 的完整性,我们确定大气湿度是主要的有害因素。温度依赖性降解速率符合阿伦尼乌斯模型,活化能为 28.5 kcal/mol,室温下的降解速率为 3.2×10(-13)/nt/s(95%置信区间:2.3-4.2/nt/s)。在这些条件下,预计每个核苷酸每 1000 个核苷酸会有 0.7-1.3 个被切割。此外,我们表明储存的 RNA 可用于进一步分析,例如逆转录定量 PCR。在模拟的几十年期间,Cq 值没有观察到明显变化。最后,我们的数据与固态 RNA 的序列无关的降解速率一致。