Abstract
MicroRNAs are a class of non-coding regulatory RNAs that can modulate development as well as alter innate antiviral defenses in plants. In this study we explored changes in Nicotiana benthamiana tobacco microRNA expression as it relates to expression of a recombinant anti-Ebola GP1 antibody. The antibody was delivered to tobacco leaves through a bacterial Agrobacterium tumefaciens “agroinfiltration” expression strategy. A multiplex microparticle-based cytometry assay tracked the expression changes of 53 host tobacco microRNAs. Our results revealed that the most abundant microRNAs in actively growing leaves corresponded to nanoparticle probes specific to nta-mir-6149 and nta-miR-168b. After agroinfiltration, probes specific for nta-mir-398, and nta-mir-482d were significantly altered in their respective expression levels, however changes were partially attributed to the infiltration broth medium used in the antibody delivery process. Confirmation of nta-mir-398 and nta-mir-482d expression changes was also verified through RT-qPCR. To our knowledge this study is the first to profile medium and Agrobacterium injection at the microRNA level through a multiplex microparticle approach.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 230-235 |
| Number of pages | 6 |
| Journal | Journal of Microbiological Methods |
| Volume | 127 |
| DOIs | |
| State | Published - Aug 1 2016 |
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Keywords
- Agrobacterium tumefaciens
- Ebola
- Nicotiana benthamiana
- Tobacco
ASJC Scopus subject areas
- Microbiology
- Molecular Biology
- Microbiology (medical)
Cite this
A cytometry microparticle platform approach for screening tobacco microRNA changes after agrobacterium delivery. / Powell, Joshua D.; Chen, Qiang; Mason, Hugh S.
In: Journal of Microbiological Methods, Vol. 127, 01.08.2016, p. 230-235.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A cytometry microparticle platform approach for screening tobacco microRNA changes after agrobacterium delivery
AU - Powell,Joshua D.
AU - Chen,Qiang
AU - Mason,Hugh S.
PY - 2016/8/1
Y1 - 2016/8/1
N2 - MicroRNAs are a class of non-coding regulatory RNAs that can modulate development as well as alter innate antiviral defenses in plants. In this study we explored changes in Nicotiana benthamiana tobacco microRNA expression as it relates to expression of a recombinant anti-Ebola GP1 antibody. The antibody was delivered to tobacco leaves through a bacterial Agrobacterium tumefaciens “agroinfiltration” expression strategy. A multiplex microparticle-based cytometry assay tracked the expression changes of 53 host tobacco microRNAs. Our results revealed that the most abundant microRNAs in actively growing leaves corresponded to nanoparticle probes specific to nta-mir-6149 and nta-miR-168b. After agroinfiltration, probes specific for nta-mir-398, and nta-mir-482d were significantly altered in their respective expression levels, however changes were partially attributed to the infiltration broth medium used in the antibody delivery process. Confirmation of nta-mir-398 and nta-mir-482d expression changes was also verified through RT-qPCR. To our knowledge this study is the first to profile medium and Agrobacterium injection at the microRNA level through a multiplex microparticle approach.
AB - MicroRNAs are a class of non-coding regulatory RNAs that can modulate development as well as alter innate antiviral defenses in plants. In this study we explored changes in Nicotiana benthamiana tobacco microRNA expression as it relates to expression of a recombinant anti-Ebola GP1 antibody. The antibody was delivered to tobacco leaves through a bacterial Agrobacterium tumefaciens “agroinfiltration” expression strategy. A multiplex microparticle-based cytometry assay tracked the expression changes of 53 host tobacco microRNAs. Our results revealed that the most abundant microRNAs in actively growing leaves corresponded to nanoparticle probes specific to nta-mir-6149 and nta-miR-168b. After agroinfiltration, probes specific for nta-mir-398, and nta-mir-482d were significantly altered in their respective expression levels, however changes were partially attributed to the infiltration broth medium used in the antibody delivery process. Confirmation of nta-mir-398 and nta-mir-482d expression changes was also verified through RT-qPCR. To our knowledge this study is the first to profile medium and Agrobacterium injection at the microRNA level through a multiplex microparticle approach.
KW - Agrobacterium tumefaciens
KW - Ebola
KW - Nicotiana benthamiana
KW - Tobacco
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UR - http://www.scopus.com/inward/citedby.url?scp=84976328955&partnerID=8YFLogxK
U2 - 10.1016/j.mimet.2016.06.023
DO - 10.1016/j.mimet.2016.06.023
M3 - Article
VL - 127
SP - 230
EP - 235
JO - Journal of Microbiological Methods
T2 - Journal of Microbiological Methods
JF - Journal of Microbiological Methods
SN - 0167-7012
ER -