Visualizing double-stranded RNA distribution and dynamics in living cells by dsRNA binding-dependent fluorescence complementation

Virology
Xiaofei ChengAiming Wang

Abstract

Double-stranded RNA (dsRNA) is an important type of RNA that plays essential roles in diverse cellular processes in eukaryotic organisms and a hallmark in infections by positive-sense RNA viruses. Currently, no in vivo technology has been developed for visualizing dsRNA in living cells. Here, we report a dsRNA binding-dependent fluorescence complementation (dRBFC) assay that can be used to efficiently monitor dsRNA distribution and dynamics in vivo. The system consists of two dsRNA-binding proteins, which are fused to the N- and C-terminal halves of the yellow fluorescent protein (YFP). Binding of the two fusion proteins to a common dsRNA brings the split YFP halves in close proximity, leading to the reconstitution of the fluorescence-competent structure and restoration of fluorescence. Using this technique, we were able to visualize the distribution and trafficking of the replicative RNA intermediates of positive-sense RNA viruses in living cells.

References

Jul 5, 1994·Proceedings of the National Academy of Sciences of the United States of America·A KumarB R Williams
Feb 11, 1994·Science·M ChalfieD C Prasher
Apr 5, 2001·Current Opinion in Biotechnology·W X Li, S W Ding
Sep 20, 2002·Journal of Virology·Frédérique Weber-LotfiLuisa Rubino
Jan 6, 2004·Nature Biotechnology·Øystein Røsok, Mouldy Sioud
Jan 31, 2006·The Plant Journal : for Cell and Molecular Biology·Keith W EarleyCraig S Pikaard
Feb 25, 2006·Cell·Shizuo AkiraOsamu Takeuchi
Dec 13, 2006·Science·Titia SijenRonald H A Plasterk
Aug 2, 2007·Immunology and Cell Biology·Alex J V Thompson, Stephen A Locarnini
Apr 17, 2008·Nature Reviews. Microbiology·Sven Miller, Jacomine Krijnse-Locker
Jul 23, 2008·The Plant Journal : for Cell and Molecular Biology·Christian WeberMarkus Fauth
Nov 5, 2008·The Plant Journal : for Cell and Molecular Biology·Jens TilsnerKarl J Oparka
Nov 7, 2008·Nature·Zachary A Bornholdt, B V Venkataram Prasad
Jan 24, 2009·The EMBO Journal·Ryuya Fukunaga, Jennifer A Doudna
Apr 1, 2009·FEBS Letters·Naoyoshi KumakuraYuichiro Watanabe
Oct 22, 2009·The Plant Journal : for Cell and Molecular Biology·Qing LuYuhai Cui
Feb 16, 2010·Trends in Plant Science·Nynne M ChristensenJens Tilsner
Jul 20, 2010·Cell Host & Microbe·Johan A den BoonPaul Ahlquist
Aug 14, 2010·Nature Reviews. Immunology·Shou-Wei Ding
Feb 8, 2011·Biological Chemistry·Yoshiko Maida, Kenkichi Masutomi
Feb 14, 2012·Trends in Microbiology·Daisy W LeungGaya K Amarasinghe
Jun 22, 2012·Journal of Virology·Romain GrangeonJean-François Laliberté
Aug 30, 2012·Nature Methods·Caroline A SchneiderKevin W Eliceiri
Sep 11, 2012·Cellular and Molecular Life Sciences : CMLS·Alys Peisley, Sun Hur
Nov 9, 2012·Analytical Chemistry·Takeaki OzawaSung Bae Kim
Sep 1, 2014·Current Opinion in Biotechnology·Anna K Rath, Andrea Rentmeister

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Citations

Feb 17, 2018·Frontiers in Plant Science·Baptiste MonsionChristophe Ritzenthaler
Sep 25, 2017·The Plant Journal : for Cell and Molecular Biology·Sang-Ho ParkAiming Wang
Sep 29, 2019·The Plant Journal : for Cell and Molecular Biology·Guanwei WuAiming Wang
Sep 3, 2020·Emerging Microbes & Infections·Sathya N Thulasi RamanXuguang Li

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