Involvement of a Rac1-Dependent Macropinocylosis Pathway in Plasmid DNA Delivery by Electrotransfection

Mao Mao; Liangli Wang; Chun-Chi Chang; Katheryn E. Rothenberg; Jianyong Huang; Yingxiao Wang; Brenton D. Hoffman; Paloma B. Liton; Fan Yuan

doi:10.1016/j.ymthe.2016.12.009

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Involvement of a Rac1-Dependent Macropinocylosis Pathway in Plasmid DNA Delivery by Electrotransfection

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Involvement of a Rac1-Dependent Macropinocylosis Pathway in Plasmid DNA Delivery by Electrotransfection

Mao Mao, Liangli Wang, Chun-Chi Chang, Katheryn E. Rothenberg, Jianyong Huang, Yingxiao Wang, Brenton D. Hoffman, Paloma B. Liton and Fan Yuan

Molecular therapy, Vol.25(3), pp.803-815

03/01/2017

DOI: 10.1016/j.ymthe.2016.12.009

PMCID: PMC5363188

PMID: 28129959

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Abstract

Electrotransfection is a widely used method for delivering genes into cells with electric pulses. Although different hypotheses have been proposed, the mechanism of electrotransfection remains controversial. Previous studies have indicated that uptake and intracellular trafficking of plasmid DNA (pDNA) are mediated by endocytic pathways, but it is still unclear which pathways are directly involved in the delivery. To this end, the present study investigated the dependence of electrotransfection on macropinocytosis. Data from the study demonstrated that electric pulses induced cell membrane ruffling and actin cytoskeleton remodeling. Using fluorescently labeled pDNA and a macropinocytosis marker (i.e., dextran), the study showed that electrotransfected pDNA co-localized with dextran in intracellular vesicles. Furthermore, electrotransfection efficiency could be decreased significantly by reducing temperature or treatment of cells with a pharmacological inhibitor of Rac1 and could be altered by changing Racl activity. Taken together, the findings suggested that electrotransfection of pDNA involved Rac1-dependent macropinocytosis.

Biotechnology & Applied Microbiology

Genetics & Heredity

Life Sciences & Biomedicine

Medicine, Research & Experimental

Research & Experimental Medicine

Science & Technology

Details

Title: Subtitle: Involvement of a Rac1-Dependent Macropinocylosis Pathway in Plasmid DNA Delivery by Electrotransfection
Creators: Mao Mao - Duke University
Liangli Wang - Duke University
Chun-Chi Chang - Duke University
Katheryn E. Rothenberg - Duke University
Jianyong Huang - Duke University
Yingxiao Wang - University of California San Diego
Brenton D. Hoffman - Duke University
Paloma B. Liton - Duke University
Fan Yuan - Duke University
Resource Type: Journal article
Publication Details: Molecular therapy, Vol.25(3), pp.803-815
DOI: 10.1016/j.ymthe.2016.12.009
PMID: 28129959
PMCID: PMC5363188
NLM abbreviation: Mol Ther
ISSN: 1525-0016
eISSN: 1525-0024
Publisher: Elsevier
Number of pages: 13
Grant note: BES-0828630 / National Science Foundation; National Science Foundation (NSF) R01EY026885 / NATIONAL EYE INSTITUTE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Eye Institute (NEI) 1344298 / Div Of Chem, Bioeng, Env, & Transp Sys; National Science Foundation (NSF); NSF - Directorate for Engineering (ENG) GM098520 / NIH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA R01GM098520 / NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS)
Language: English
Date published: 03/01/2017
Academic Unit: Biology
Record Identifier: 9984696710902771

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