RNA viral vectors for improved Agrobacterium-mediated transient expression of heterologous proteins in Nicotiana benthamiana cell suspensions and hairy roots.

BMC Biotechnology
Jeffrey S Larsen, Wayne R Curtis

Abstract

Plant cell suspensions and hairy root cultures represent scalable protein expression platforms. Low protein product titers have thus far limited the application of transient protein expression in these hosts. The objective of this work was to overcome this limitation by harnessing A. tumefaciens to deliver replicating and non-replicating RNA viral vectors in plant tissue co-cultures. Replicating vectors derived from Potato virus X (PVX) and Tobacco rattle virus (TRV) were modified to contain the reporter gene β-glucuronidase (GUS) with a plant intron to prevent bacterial expression. In cell suspensions, a minimal PVX vector retaining only the viral RNA polymerase gene yielded 6.6-fold more GUS than an analogous full-length PVX vector. Transient co-expression of the minimal PVX vector with P19 of Tomato bushy stunt virus or HC-Pro of Tobacco etch virus to suppress post-transcriptional gene silencing increased GUS expression by 44 and 83%, respectively. A non-replicating vector containing a leader sequence from Cowpea mosaic virus (CPMV-HT) modified for enhanced translation led to 70% higher transient GUS expression than a control treatment. In hairy roots, a TRV vector capable of systemic movement increased GUS accumulation by 1...Continue Reading

References

Apr 1, 1968·Experimental Cell Research·O L GamborgK Ojima
Apr 21, 1999·Current Opinion in Biotechnology·J V Shanks, J Morgan
Oct 6, 1999·Current Opinion in Plant Biology·K M Crawford, P C Zambryski
Dec 2, 1999·The Journal of General Virology·S A MacFarlane
Jan 29, 2000·Virology·S A MacFarlane, A H Popovich
Feb 7, 2001·Current Opinion in Biotechnology·P M Doran
Feb 13, 2001·FEBS Letters·F GilJ M Escribano
May 25, 2002·The Plant Journal : for Cell and Molecular Biology·Yule LiuS P Dinesh-Kumar
Mar 1, 2003·The Plant Journal : for Cell and Molecular Biology·Olivier VoinnetDavid Baulcombe
Apr 8, 2004·Virus Research·Braden M RothVicki B Vance
Jun 5, 2004·Biotechnology Progress·Jason I CollensWayne R Curtis
Sep 28, 2004·Biotechnology and Bioengineering·Divakar Ramakrishnan, Wayne R Curtis
Dec 14, 2004·Plant Physiology·Tracy ValentineChristophe Lacomme
Aug 23, 2005·Journal of Virological Methods·I A MechtcheriakovaG P Lomonossoff
Oct 8, 2005·Environmental Biosafety Research·Ulrike Manske, Joachim Schiemann
Jul 18, 2006·Trends in Biotechnology·Pauline M Doran
Sep 16, 2006·Proceedings of the National Academy of Sciences of the United States of America·Anatoli GiritchYuri Gleba
Sep 19, 2006·Biochemistry. Biokhimii︠a︡·T V KomarovaJ G Atabekov
Feb 29, 2008·Biotechnology and Bioengineering·Marina SkarjinskaiaVidadi Yusibov
Feb 3, 2009·Methods in Molecular Biology·Jose M Escribano, Daniel M Perez-Filgueira
Mar 25, 2009·Biotechnology and Bioengineering·Zhong HuangHugh Mason
Jan 1, 2007·Influenza and Other Respiratory Viruses·Konstantin MusiychukVidadi Yusibov
Jul 25, 2009·Plant Biotechnology Journal·Frank SainsburyGeorge P Lomonossoff
Dec 4, 2009·Applied Microbiology and Biotechnology·Pamela J WeathersJianfeng Xu
Jan 7, 2010·Plant Biotechnology Journal·Charles GouletDominique Michaud
Mar 1, 2003·Molecular Plant Pathology·Jeffrey S BattenCynthia Hemenway
Dec 18, 2010·BMC Biotechnology·Zun Liu, Christopher M Kearney

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Citations

Mar 13, 2013·Plant Molecular Biology·Hadrien Peyret, George P Lomonossoff
Dec 4, 2013·Plant Physiology·Aihua ShaYule Liu
Apr 17, 2015·Plant Biotechnology Journal·Thangarasu MuthamilselvanYau-Heiu Hsu
Jun 16, 2015·Plant Biotechnology Journal·Hadrien Peyret, George P Lomonossoff
Oct 19, 2014·Biotechnology Letters·Shah FahadJianliang Huang
Jul 22, 2016·Plant Biotechnology Journal·Naila Shahid, Henry Daniell
May 7, 2016·Chemical Society Reviews·Amy M Wen, Nicole F Steinmetz
Apr 17, 2018·International Journal of Molecular Sciences·Sara C SukenikKaren A McDonald
Oct 9, 2020·The Plant Journal : for Cell and Molecular Biology·Poonam SinghMariam Gaid

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Datasets Mentioned

BETA
AY297843

Methods Mentioned

BETA
transgenic
PCR
restriction digest
transfection

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