Highly Efficient and Robust π-FISH Rainbow for Multiplexed in Situ Detection of Diverse Biomolecules

Overview

Journal Nat Commun

Specialty Biology

Date 2023 Jan 27

PMID 36707540

Authors

Yingfeng Tao

Xiaoliu Zhou

Leqiang Sun

Da Lin

Huaiyuan Cai

Xi Chen

Wei Zhou

Bing Yang

Zhe Hu

Jing Yu

Jing Zhang

Xiaoqing Yang

Fang Yang

Bang Shen

Wenbao Qi

Zhenfang Fu

Jinxia Dai

Gang Cao

Affiliations

Soon will be listed here.

Abstract

In the unprecedented single-cell sequencing and spatial multiomics era of biology, fluorescence in situ hybridization (FISH) technologies with higher sensitivity and robustness, especially for detecting short RNAs and other biomolecules, are greatly desired. Here, we develop the robust multiplex π-FISH rainbow method to detect diverse biomolecules (DNA, RNA, proteins, and neurotransmitters) individually or simultaneously with high efficiency. This versatile method is successfully applied to detect gene expression in different species, from microorganisms to plants and animals. Furthermore, we delineate the landscape of diverse neuron subclusters by decoding the spatial distribution of 21 marker genes via only two rounds of hybridization. Significantly, we combine π-FISH rainbow with hybridization chain reaction to develop π-FISH+ technology for short nucleic acid fragments, such as microRNA and prostate cancer anti-androgen therapy-resistant marker ARV7 splicing variant in circulating tumour cells from patients. Our study provides a robust biomolecule in situ detection technology for spatial multiomics investigation and clinical diagnosis.

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