title: Visualization of NRAS RNA G-Quadruplex Structures in Cells with an Engineered Fluorogenic Hybridization Probe.
abstract: The RNA G-quadruplex is an important secondary structure formed by guanine-rich RNA sequences. However, its folding studies have mainly been studied in vitro. Accurate identification of RNA G-quadruplex formation within a sequence of interest remains difficult in cells. Herein, and based on the guanine-rich sequence in the 5'-UTR of NRAS mRNA, we designed and synthesized the first G-quadruplex-triggered fluorogenic hybridization (GTFH) probe, ISCH-nras1, for the unique visualization of the G-quadruplexes that form in this region. ISCH-nras1 is made up of two parts: The first is a fluorescent light-up moiety specific to G-quadruplex structures, and the second is a DNA molecule that can hybridize with a sequence that is adjacent to the guanine-rich sequence in the NRAS mRNA 5'-UTR. Further evaluation studies indicated that ISCH-nras1 could directly and precisely detect the targeted NRAS RNA G-quadruplex structures, both in vitro and in cells. Thus, this GTFH probe was a useful tool for directly investigating the folding of G-quadruplex structures within an RNA of interest and represents a new direction for the design of smart RNA G-quadruplex probes.
authors: School of Pharmaceutical Sciences, Sun Yat-sen University , Guangzhou 510006, China.
authors: Shuo-Bin
authors: SB
authors: Chen
authors: School of Pharmaceutical Sciences, Sun Yat-sen University , Guangzhou 510006, China.
authors: Ming-Hao
authors: MH
authors: Hu
authors: School of Pharmaceutical Sciences, Sun Yat-sen University , Guangzhou 510006, China.
authors: Guo-Cai
authors: GC
authors: Liu
authors: Guangzhou Institute of Geochemistry, Chinese Academy of Sciences , Guangzhou 510640, China.
authors: Jin
authors: J
authors: Wang
authors: School of Pharmaceutical Sciences, Sun Yat-sen University , Guangzhou 510006, China.
authors: Tian-Miao
authors: TM
authors: Ou
authors: School of Pharmaceutical Sciences, Sun Yat-sen University , Guangzhou 510006, China.
authors: Lian-Quan
authors: LQ
authors: Gu
authors: School of Pharmaceutical Sciences, Sun Yat-sen University , Guangzhou 510006, China.
authors: Zhi-Shu
authors: ZS
authors: Huang
authors: School of Pharmaceutical Sciences, Sun Yat-sen University , Guangzhou 510006, China.
authors: Jia-Heng
authors: JH
authors: Tan
mesh_terms: D001483:Base Sequence; D004738:Engineering; D005456:Fluorescent Dyes / Q000378:metabolism*; D054856:G-Quadruplexes; D020558:GTP Phosphohydrolases / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D012313:RNA; D020558:GTP Phosphohydrolases