A mechanistic investigation exploring the differential transfection efficiencies between the easy-to-transfect SK-BR3 and difficult-to-transfect CT26 cell lines

Simple item page
dc.citation.articleNumber36en_US
dc.citation.issueNumber1en_US
dc.citation.journalTitleJournal of Nanobiotechnologyen_US
dc.citation.volumeNumber15en_US
dc.contributor.authorFigueroa, Elizabethen_US
dc.contributor.authorBugga, Pallavien_US
dc.contributor.authorAsthana, Vishwaratnen_US
dc.contributor.authorChen, Allen L.en_US
dc.contributor.authorStephen Yan, J.en_US
dc.contributor.authorEvans, Emily R.en_US
dc.contributor.authorDrezek, Rebekah A.en_US
dc.date.accessioned2017-05-02T16:02:30Zen_US
dc.date.available2017-05-02T16:02:30Zen_US
dc.date.issued2017en_US
dc.date.updated2017-05-02T16:02:30Zen_US
dc.description.abstractAbstract Background Gold–polyamidoamine (AuPAMAM) has previously been shown to successfully transfect cells with high efficiency. However, we have observed that certain cell types are more amenable to Au–PAMAM transfection than others. Here we utilized two representative cell lines—a “difficult to transfect” CT26 cell line and an “easy to transfect” SK-BR3 cell line—and attempted to determine the underlying mechanism for differential transfection in both cell types. Using a commonly established poly-cationic polymer similar to PAMAM (polyethyleneimine, or PEI), we additionally sought to quantify the relative transfection efficiencies of each vector in CT26 and SK-BR3 cells, in the hopes of elucidating any mechanistic differences that may exist between the two transfection vectors. Results A comparative time course analysis of green fluorescent protein reporter-gene expression and DNA uptake was conducted to quantitatively compare PEI- and AuPAMAM-mediated transfection in CT26 and SK-BR3, while flow cytometry and confocal microscopy were used to determine the contribution of cellular uptake, endosomal escape, and cytoplasmic transport to the overall gene delivery process. Results from the time course analysis and flow cytometry studies revealed that initial complex uptake and cytoplasmic trafficking to the nucleus are likely the two main factors limiting CT26 transfectability. Conclusions The cell type-dependent uptake and intracellular transport mechanisms impacting gene therapy remain largely unexplored and present a major hurdle in the application-specific design and efficiency of gene delivery vectors. This systematic investigation offers insights into the intracellular mechanistic processes that may account for cell-to-cell differences, as well as vector-to-vector differences, in gene transfectability.en_US
dc.identifier.citationFigueroa, Elizabeth, Bugga, Pallavi, Asthana, Vishwaratn, et al.. "A mechanistic investigation exploring the differential transfection efficiencies between the easy-to-transfect SK-BR3 and difficult-to-transfect CT26 cell lines." <i>Journal of Nanobiotechnology,</i> 15, no. 1 (2017) BioMed Central: http://dx.doi.org/10.1186/s12951-017-0271-8.en_US
dc.identifier.doihttp://dx.doi.org/10.1186/s12951-017-0271-8en_US
dc.identifier.urihttps://hdl.handle.net/1911/94110en_US
dc.language.isoengen_US
dc.publisherBioMed Centralen_US
dc.rightsThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.en_US
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en_US
dc.titleA mechanistic investigation exploring the differential transfection efficiencies between the easy-to-transfect SK-BR3 and difficult-to-transfect CT26 cell linesen_US
dc.typeJournal articleen_US
dc.type.dcmiTexten_US
dc.type.publicationpublisher versionen_US
local.sword.agentBioMed Centralen_US
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
12951_2017_Article_271.pdf
Size:
2.99 MB
Format:
Adobe Portable Document Format
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.61 KB
Format:
Item-specific license agreed upon to submission
Description:
Download
Collections
Faculty Publications
Bioengineering Publications