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Analysis and Identification of Tumorigenic Targets of MicroRNA in Cancer Cells by Photoreactive Chemical Probes

School of Chemistry & Chemical Engineering, Inner Mongolia Key Laboratory of Mongolian Medicinal Chemistry, Inner Mongolia University, 235 West College Road, Hohhot, Inner Mongolia 010020, China
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Int. J. Mol. Sci. 2020, 21(4), 1545; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21041545
Received: 19 January 2020 / Revised: 6 February 2020 / Accepted: 18 February 2020 / Published: 24 February 2020
(This article belongs to the Section Molecular Biology)
Photoactive RNA probes have unique advantages in the identification of microRNA (miR) targets due to their ability for efficient conjugation to the target sequences by covalent crosslinking, providing stable miR-mRNA complexes for further analysis. Here, we report a highly efficient and straightforward method for miR target identification that is based on photo-reactive chemical probes and RNA-seq technology (denotes PCP-Seq). UV reactive probes were prepared by incorporating psoralen in the specific position of the seed sequence of miR. Cancer cells that were transfected with the miR probes were treated with UV, following the isolation of poly(A) RNA and sequencing of the transcriptome. Quantitative analysis of RNA-seq reads and subsequent validation by qPCR, dual luciferase assay as well as western blotting confirmed that PCP-Seq could highly efficiently identify multiple targets of different miRs in the lung cancer cell line, such as targets PTTG1 and PTGR1 of miR-29a and ILF2 of miR-34a. Collectively, our data showed that PCP-Seq is a robust strategy for miR targets identification, and unique in the identification of the targets that escape degradation by miRISC and maintain normal cellular level, although their translation is repressed. View Full-Text
Keywords: chemical probe; RNA interference; microRNA target; RNA-seq; tumorigenic gene chemical probe; RNA interference; microRNA target; RNA-seq; tumorigenic gene
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MDPI and ACS Style

Su, Z.; Ganbold, T.; Baigude, H. Analysis and Identification of Tumorigenic Targets of MicroRNA in Cancer Cells by Photoreactive Chemical Probes. Int. J. Mol. Sci. 2020, 21, 1545. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21041545

AMA Style

Su Z, Ganbold T, Baigude H. Analysis and Identification of Tumorigenic Targets of MicroRNA in Cancer Cells by Photoreactive Chemical Probes. International Journal of Molecular Sciences. 2020; 21(4):1545. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21041545

Chicago/Turabian Style

Su, Zhiyu, Tsogzolmaa Ganbold, and Huricha Baigude. 2020. "Analysis and Identification of Tumorigenic Targets of MicroRNA in Cancer Cells by Photoreactive Chemical Probes" International Journal of Molecular Sciences 21, no. 4: 1545. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21041545

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