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Intracellular Availability of pDNA and mRNA after Transfection: A Comparative Study among Polyplexes, Lipoplexes, and Lipopolyplexes
journal contribution
posted on 2016-08-03, 00:00 authored by Cristine Gonçalves, Sohail Akhter, Chantal Pichon, Patrick MidouxIntracellular availability
of nucleic acids from synthetic vectors
is critical and directly influences the transfection efficiency (TE).
Herein, we evaluated the TE of polymer- and lipid-based nanoplexes
(polyplexes, lipoplexes and lipopolyplexes) of EGFP-encoding mRNA
and pDNA. To determine the translation and transcription efficiency
of each nucleic acid nanoplex, in vitro expression
was measured in HEK293T7 cells that permit gene expression in the
cytoplasmic region. Globally, mRNA transfection profile was well corroborative
with cytoplasmic transfection of pT7-pDNA as well as with nuclear
transfection of pCMV-DNA. Irrespective of the nucleic acid, high TE
was observed with histidinylated l-polyethylenimine (His-lPEI) polyplexes
and dioleyl succinyl paromomycin/O,O-dioleyl-N-histamine phosphoramidate (DOPS/MM27)
lipoplexes. Moreover, His-lPEI polyplexes yielded higher in
vitro expression of EGFP for pDNA than for mRNA. Furthermore,
a significant enhancement in the TE in the presence of an excess of
His-lPEI was observed indicating that this polymer promotes cytosolic
delivery. Compared to other nanoplexes, His-lPEI polyplex showed high
intracellular availability of DNA and mRNA along with low cytotoxicity,
owing to its rapid (complete or partial) unpacking in the cytosol
and/or endosomes. This study gives an insight that, whether with mRNA
or pDNA, enhancing nanoplex unpacking in the endosomes and cytosol
would improve the delivery of nucleic acid in the cytosol and particularly
in the case of pDNA where a sufficient available amount of pDNA in
the cytoplasm would definitely improve its transport toward the nucleus.
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Keywords
histamine phosphoramidateLipopolyplexes Intracellular availabilityacid nanoplexTEintracellular availabilitycytoplasmic transfectionIntracellular Availabilitygene expressiontranscription efficiencyDNAHEK 293T cellsEGFPcytoplasmic regionmRNA transfection profiletransfection efficiencypDNAcytosolic deliveryComparative StudyDOPS
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