RNA G-quadruplex formation in defined sequence in living cells detected by bimolecular fluorescence complementation
| Autor: | Ke-wei Zheng, Quan Chen, Zheng Tan, Yi-de He, Yu-hua Hao, Hong-he Liu |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Riboswitch RNA Nuclease protection assay General Chemistry Biology Molecular biology Post-transcriptional modification Cell biology Chemistry 03 medical and health sciences Bimolecular fluorescence complementation chemistry.chemical_compound 030104 developmental biology chemistry Regulatory sequence RNA editing heterocyclic compounds DNA |
| Zdroj: | Chemical Science |
| ISSN: | 2041-6539 2041-6520 |
| DOI: | 10.1039/c5sc03946k |
| Popis: | RNA G-quadruplex formation in a defined sequence in living bacterial cells is detected by bimolecular fluorescence complementation with sequence and structure specificity. G-quadruplexes are implicated in many essential cellular processes and sequences with potential to form a G-quadruplex are widely present in DNA and RNA. However, it is difficult to know whether a sequence of interest naturally forms a G-quadruplex in living cells. Here we report the detection of a G-quadruplex in defined RNA sequences in living cells in a natural intracellular environment. A G-quadruplex forming sequence in a RNA transcript is tagged at proximity with an aptamer. The two structures are recognized respectively by two probe proteins each of which is fused with a split half of enhanced green fluorescent protein (eGFP). Simultaneous binding of the two proteins to RNA brings the two halves of eGFP into proximity, permitting them to reconstitute into a functional eGFP that emits fluorescence to signal the formation of a G-quadruplex in RNA. We show that a G-quadruplex can form in RNA and can be detected with sequence and structure specificity under both in vitro and in vivo conditions. The results, therefore, provide direct evidence for the formation of RNA G-quadruplexes in live cells and the method provides a useful tool to validate G-quadruplex formation in a specific sequence under a natural cellular condition. |
| Databáze: | OpenAIRE |
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