利用 sRNA-FISH 技术在植物中进行小分子 RNA 的荧光原位检测。

Fluorescent In Situ Detection of Small RNAs in Plants Using sRNA-FISH.

机构信息

Department of Plant and Soil Sciences, University of Delaware, Newark, DE, USA.

Bio-Imaging Center, Delaware Biotechnology Institute, University of Delaware, Newark, DE, USA.

出版信息

Methods Mol Biol. 2024;2784:101-111. doi: 10.1007/978-1-0716-3766-1_7.

DOI:10.1007/978-1-0716-3766-1_7

PMID:38502481

Abstract

Plant small RNAs are 21-24 nucleotide, noncoding RNAs that function as regulators in plant growth and development. Colorimetric detection of plant small RNAs was made possible with the introduction of locked nucleic acid probes. However, fluorescent detection of plant small RNAs has been challenging due to the high autofluorescence from plant tissue. Here we report a fluorescent in situ detection method for plant small RNAs. This method can be applied to most plant samples and tissue types and also can be adapted for single-molecule detection of small RNAs with super-resolution microscopy.

摘要

植物小 RNA 是 21-24 个核苷酸的非编码 RNA，在植物生长和发育中作为调节剂发挥作用。引入锁核酸探针使得植物小 RNA 的比色检测成为可能。然而，由于植物组织的高自发荧光，植物小 RNA 的荧光检测一直具有挑战性。本文报道了一种用于植物小 RNA 的荧光原位检测方法。该方法可应用于大多数植物样本和组织类型，也可适应超分辨率显微镜对小 RNA 的单分子检测进行改编。