PT  - JOURNAL ARTICLE
AU  - Hirokazu Takahashi
AU  - Kyohei Horio
AU  - Setsu Kato
AU  - Toshiro Kobori
AU  - Kenshi Watanabe
AU  - Tsunehiro Aki
AU  - Yutaka Nakashimada
AU  - Yoshiko Okamura
TI  - Direct detection of mRNA expression in microbial cells by fluorescence <em>in situ</em> hybridization using RNase H-assisted rolling circle amplification
AID  - 10.1101/729616
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 729616
4099  - http://biorxiv.org/content/early/2019/11/21/729616.short
4100  - http://biorxiv.org/content/early/2019/11/21/729616.full
AB  - Meta-analyses using next generation sequencing is a powerful strategy for studying microbiota; however, it cannot clarify the role of individual microbes within microbiota. To know which cell expresses what gene is important for elucidation of the individual cell’s function in microbiota. In this report, we developed novel fluorescence in situ hybridization (FISH) procedure using RNase-H-assisted rolling circle amplification to visualize mRNA of interest in microbial cells without reverse transcription. Our results show that this method is applicable to both gram-negative and gram-positive microbes without any noise from DNA, and it is possible to visualize the target mRNA expression directly at the single-cell level. Therefore, our procedure, when combined with data of meta-analyses, can help to understand the role of individual microbes in the microbiota.