从未经细胞分选的冰冻组织样品中分离细胞类型特异性 RNA。
Isolate Cell-Type-Specific RNAs from Snap-Frozen Heterogeneous Tissue Samples without Cell Sorting.
机构信息
Department of Anatomy, Physiology and Pharmacology, College of Veterinary Medicine, Auburn University;
Department of Anatomy, Physiology and Pharmacology, College of Veterinary Medicine, Auburn University.
出版信息
J Vis Exp. 2021 Dec 8(178). doi: 10.3791/63143.
Cellular heterogeneity poses challenges to understanding the function of complex tissues at a transcriptome level. Using cell-type-specific RNAs avoids potential pitfalls caused by the heterogeneity of tissues and unleashes the powerful transcriptome analysis. The protocol described here demonstrates how to use the Translating Ribosome Affinity Purification (TRAP) method to isolate ribosome-bound RNAs from a small amount of EGFP-expressing cells in a complex tissue without cell sorting. This protocol is suitable for isolating cell-type-specific RNAs using the recently available NuTRAP mouse model and could also be used to isolate RNAs from any EGFP-expressing cells.
细胞异质性给在转录组水平理解复杂组织的功能带来了挑战。使用细胞类型特异性 RNA 可以避免组织异质性带来的潜在陷阱，并释放强大的转录组分析能力。本方案描述了如何在没有细胞分选的情况下，从小鼠复杂组织中少量表达 EGFP 的细胞中，使用核糖体亲和纯化（TRAP）方法分离核糖体结合的 RNA。该方案适用于使用最近可用的 NuTRAP 小鼠模型分离细胞类型特异性 RNA，也可用于分离任何表达 EGFP 的细胞的 RNA。
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