MARIS: Method for Analyzing RNA following Intracellular Sorting

Transcriptional profiling is a key technique in the study of cell biology that is limited by the availability of reagents to uniquely identify specific cell types and isolate high quality RNA from them. We report a Method for Analyzing RNA following Intracellular Sorting (MARIS) that generates high quality RNA for transcriptome profiling following cellular fixation, intracellular immunofluorescent staining and FACS. MARIS can therefore be used to isolate high quality RNA from many otherwise inaccessible cell types simply based on immunofluorescent tagging of unique intracellular proteins. As proof of principle, we isolate RNA from sorted human embryonic stem cell-derived insulin-expressing cells as well as adult human β cells. MARIS is a basic molecular biology technique that could be used across several biological disciplines.

转录组分析（Transcriptional profiling）是细胞生物学研究中的关键技术，但其应用受制于试剂的可获得性：此类试剂需能够特异性识别特定细胞类型，并从中分离高质量RNA。本研究报道了一种名为细胞内分选后RNA分析（MARIS, Method for Analyzing RNA following Intracellular Sorting）的方法，该方法可在细胞固定、胞内免疫荧光染色及流式荧光激活细胞分选术（Fluorescence-Activated Cell Sorting，FACS）分选后，获取高质量RNA以用于转录组分析。因此，MARIS技术仅需通过对特异性胞内蛋白进行免疫荧光标记，即可从诸多原本难以获取的细胞类型中分离得到高质量RNA。作为原理验证，我们分别从分选获得的人胚胎干细胞衍生的胰岛素表达细胞以及成人人类β细胞中分离了RNA。MARIS作为一项基础分子生物学技术，可应用于多个生物学科领域。