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

Publication ,  Journal Article
Mao, M; Wang, L; Chang, C-C; Rothenberg, KE; Huang, J; Wang, Y; Hoffman, BD; Liton, PB; Yuan, F
Published in: Mol Ther
March 1, 2017

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 Rac1 activity. Taken together, the findings suggested that electrotransfection of pDNA involved Rac1-dependent macropinocytosis.

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Published In

Mol Ther

DOI

EISSN

1525-0024

Publication Date

March 1, 2017

Volume

25

Issue

3

Start / End Page

803 / 815

Location

United States

Related Subject Headings

  • rac1 GTP-Binding Protein
  • Transfection
  • Plasmids
  • Pinocytosis
  • Microscopy, Fluorescence
  • Mice
  • Humans
  • Gene Transfer Techniques
  • Gene Expression
  • Endocytosis
 

Citation

APA
Chicago
ICMJE
MLA
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Mao, M., Wang, L., Chang, C.-C., Rothenberg, K. E., Huang, J., Wang, Y., … Yuan, F. (2017). Involvement of a Rac1-Dependent Macropinocytosis Pathway in Plasmid DNA Delivery by Electrotransfection. Mol Ther, 25(3), 803–815. https://doi.org/10.1016/j.ymthe.2016.12.009
Mao, Mao, Liangli Wang, Chun-Chi Chang, Katheryn E. Rothenberg, Jianyong Huang, Yingxiao Wang, Brenton D. Hoffman, Paloma B. Liton, and Fan Yuan. “Involvement of a Rac1-Dependent Macropinocytosis Pathway in Plasmid DNA Delivery by Electrotransfection.Mol Ther 25, no. 3 (March 1, 2017): 803–15. https://doi.org/10.1016/j.ymthe.2016.12.009.
Mao M, Wang L, Chang C-C, Rothenberg KE, Huang J, Wang Y, et al. Involvement of a Rac1-Dependent Macropinocytosis Pathway in Plasmid DNA Delivery by Electrotransfection. Mol Ther. 2017 Mar 1;25(3):803–15.
Mao, Mao, et al. “Involvement of a Rac1-Dependent Macropinocytosis Pathway in Plasmid DNA Delivery by Electrotransfection.Mol Ther, vol. 25, no. 3, Mar. 2017, pp. 803–15. Pubmed, doi:10.1016/j.ymthe.2016.12.009.
Mao M, Wang L, Chang C-C, Rothenberg KE, Huang J, Wang Y, Hoffman BD, Liton PB, Yuan F. Involvement of a Rac1-Dependent Macropinocytosis Pathway in Plasmid DNA Delivery by Electrotransfection. Mol Ther. 2017 Mar 1;25(3):803–815.

Published In

Mol Ther

DOI

EISSN

1525-0024

Publication Date

March 1, 2017

Volume

25

Issue

3

Start / End Page

803 / 815

Location

United States

Related Subject Headings

  • rac1 GTP-Binding Protein
  • Transfection
  • Plasmids
  • Pinocytosis
  • Microscopy, Fluorescence
  • Mice
  • Humans
  • Gene Transfer Techniques
  • Gene Expression
  • Endocytosis