L-DNA 标记荧光杂交技术用于单细胞中 RNA 的高灵敏度成像。

L-DNA-tagged fluorescence hybridization for highly sensitive imaging of RNAs in single cells.

机构信息

Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.

Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.

出版信息

Org Biomol Chem. 2020 Oct 21;18(40):8084-8088. doi: 10.1039/d0ob01635g.

DOI:10.1039/d0ob01635g

PMID:33001106

Abstract

We report an effective fluorescence in situ hybridization strategy, named l-DNA tagged FISH (LT-FISH), for highly sensitive RNA detection in fixed cultured cells. LT-FISH includes two-step hybridization processes with a l-d chimera oligonucleotide probe and a fluorescence-labeled PCR product tethering a l-DNA tag. The degree of fluorescence enhancement, depending on the length of PCR products, was up to 14-fold when the 606 bp product was used. Endogenous mRNA and miRNA in cancer cells were visualized by utilizing this l-DNA-mediated signal amplification technique.

摘要

我们报告了一种有效的荧光原位杂交策略，称为 l-DNA 标记 FISH（LT-FISH），用于在固定培养细胞中高度灵敏地检测 RNA。LT-FISH 包括两步杂交过程，使用 l-d 嵌合体寡核苷酸探针和连接 l-DNA 标签的荧光标记 PCR 产物。当使用 606 bp 产物时，荧光增强程度高达 14 倍。利用这种 l-DNA 介导的信号放大技术，可以可视化癌细胞中的内源性 mRNA 和 miRNA。