Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Jan;1(1):e28.
doi: 10.1002/cpz1.28.

Observation of Z-DNA Structure via the Synthesis of Oligonucleotide DNA Containing 8-Trifluoromethyl-2-Deoxyguanosine

Hong-Liang Bao  1 , Yan Xu  1
Affiliations

Observation of Z-DNA Structure via the Synthesis of Oligonucleotide DNA Containing 8-Trifluoromethyl-2-Deoxyguanosine

Hong-Liang Bao et al. Curr Protoc. 2021 Jan.

Erratum in

Abstract

This article contains detailed synthetic protocols for the preparation of DNA oligonucleotides containing 8-trifluoromethyl-2'-deoxyguanosine (CF3 dG) and their application to observe Z-DNA structure in vitro and in living HeLa cells. First, using a catalytic system consisting of FeSO4 , H2 SO4 , and H2 O2 in DMSO, we achieved a one-step synthesis of CF3 dG through a radical reaction between deoxyguanosine (dG) and CF3 I, with a yield of 45%. We then obtained the 3'-phosphoramidite of CF3 dG through a routine three-step procedure. Next, we employed the CF3 dG phosphoramidite monomer in the synthesis of oligonucleotides on a solid-phase DNA synthesizer. Finally, we used the CF3 dG-modified DNA oligonucleotides to observe Z-DNA structure in vitro and in living HeLa cells through 19 F NMR spectroscopy. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Synthesis of CF3 dG phosphoramidites Basic Protocol 2: Preparation of CF3 dG-modified DNA oligonucleotides Basic Protocol 3: Evaluation of CF3 dG stabilization of Z-DNA structure by CD spectroscopy Basic Protocol 4: Investigation of Z-DNA structure in vitro and in HeLa cells with CF3 dG-modified DNA oligonucleotides and 19 F NMR spectroscopy.

Keywords: 19F NMR spectroscopy; 8-trifluoromethyl-2′-deoxyguanosine; Z-DNA; in-cell NMR.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

  • ADAR1 masks the cancer immunotherapeutic promise of ZBP1-driven necroptosis.
    Zhang T, Yin C, Fedorov A, Qiao L, Bao H, Beknazarov N, Wang S, Gautam A, Williams RM, Crawford JC, Peri S, Studitsky V, Beg AA, Thomas PG, Walkley C, Xu Y, Poptsova M, Herbert A, Balachandran S. Zhang T, et al. Nature. 2022 Jun;606(7914):594-602. doi: 10.1038/s41586-022-04753-7. Epub 2022 May 25. Nature. 2022. PMID: 35614224 Free PMC article.

References

Literature Cited

    1. Balasubramaniyam, T., Ishizuka, T., Xiao, C. D., Bao, H. L. & Xu, Y. (2019). 2′-O-Methyl-8-methylguanosine as a Z-form RNA stabilizer for structural and functional study of Z-RNA. Molecules, 23, 2572. doi: 10.3390/molecules23102572.
    1. Bao, H. L., Masuzawa, T., Oyoshi, T., & Xu, Y. (2020). Oligonucleotides DNA containing 8-trifluoromethyl-2′-deoxyguanosine for observing Z-DNA structure. Nucleic Acids Research, 48, 7041-7051. doi: 10.1093/nar/gkaa505.
    1. Bao, H. L., Liu, H. S., & Xu, Y. (2019). Hybrid-type and two-tetrad antiparallel telomere DNA G-quadruplex structures in living human cells. Nucleic Acids Research, 47, 4940-4947. doi: 10.1093/nar/gkz276.
    1. Bao, H. L., & Xu, Y. (2018). Investigation of higher-order RNA G-quadruplex structures in vitro and in living cells by 19F NMR spectroscopy. Nature Protocols, 13, 652-665. doi: 10.1038/nprot.2017.156.
    1. Bao, H. L., Ishizuka, T., Sakamoto, T., Fujimoto, K., Uechi, T., Kenmochi, N., & Xu, Y. (2017). Characterization of human telomere RNA G-quadruplex structures in vitro and in living cells using 19F NMR spectroscopy. Nucleic Acids Research, 45, 5501-5511. doi: 10.1093/nar/gkx109.

LinkOut - more resources