核酸
DNA
核糖核酸
锁核酸
寡核苷酸
生物
杂交探针
底漆(化妆品)
化学
计算生物学
分子生物学
遗传学
基因
有机化学
作者
Jocelyn Y. Kishi,Sylvain W. Lapan,Brian J. Beliveau,Emma R. West,Allen Zhu,Hiroshi Sasaki,Sinem K. Saka,Yu Wang,Constance L. Cepko,Peng Yin
出处
期刊:Nature Methods
[Springer Nature]
日期:2019-05-20
卷期号:16 (6): 533-544
被引量:265
标识
DOI:10.1038/s41592-019-0404-0
摘要
Fluorescence in situ hybridization (FISH) reveals the abundance and positioning of nucleic acid sequences in fixed samples. Despite recent advances in multiplexed amplification of FISH signals, it remains challenging to achieve high levels of simultaneous amplification and sequential detection with high sampling efficiency and simple workflows. Here we introduce signal amplification by exchange reaction (SABER), which endows oligonucleotide-based FISH probes with long, single-stranded DNA concatemers that aggregate a multitude of short complementary fluorescent imager strands. We show that SABER amplified RNA and DNA FISH signals (5- to 450-fold) in fixed cells and tissues. We also applied 17 orthogonal amplifiers against chromosomal targets simultaneously and detected mRNAs with high efficiency. We then used 10-plex SABER-FISH to identify in vivo introduced enhancers with cell-type-specific activity in the mouse retina. SABER represents a simple and versatile molecular toolkit for rapid and cost-effective multiplexed imaging of nucleic acid targets. Using primer-exchange reactions, SABER extends FISH probes with repetitive sequences that can accommodate multiple fluorescent imager strands, resulting in up to 450-fold signal amplification. SABER is showcased in DNA and RNA FISH experiments across a range of complex biological samples.
科研通智能强力驱动
Strongly Powered by AbleSci AI