title: Intracellular Availability of pDNA and mRNA after Transfection: A Comparative Study among Polyplexes, Lipoplexes, and Lipopolyplexes.
abstract: Intracellular 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.
authors.affiliation: Centre de Biophysique Moléculaire, CNRS UPR4301 , rue Charles Sadron CS 80054, F-45071 Orléans Cedex 02, France.|Université d'Orléans , Orléans, France.
authors.forename: Cristine
authors.initials: C
authors.lastname: Gonçalves
authors.affiliation: Centre de Biophysique Moléculaire, CNRS UPR4301 , rue Charles Sadron CS 80054, F-45071 Orléans Cedex 02, France.|Université d'Orléans , Orléans, France.|Le Studium Loire Valley Institute for Advanced Studies , Centre-Val de Loire région, France.
authors.forename: Sohail
authors.initials: S
authors.lastname: Akhter
authors.affiliation: Centre de Biophysique Moléculaire, CNRS UPR4301 , rue Charles Sadron CS 80054, F-45071 Orléans Cedex 02, France.|Université d'Orléans , Orléans, France.
authors.forename: Chantal
authors.initials: C
authors.lastname: Pichon
authors.affiliation: Centre de Biophysique Moléculaire, CNRS UPR4301 , rue Charles Sadron CS 80054, F-45071 Orléans Cedex 02, France.|Université d'Orléans , Orléans, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Midoux
mesh_terms: D002460:Cell Line; D003593:Cytoplasm / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D011992:Endosomes / Q000378:metabolism; D018014:Gene Transfer Techniques; D006801:Humans; D008055:Lipids / Q000737:chemistry; D010316:Particle Size; D010957:Plasmids / Q000737:chemistry*; D011094:Polyethyleneimine / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
chemical_list: D008055:Lipids; D011108:Polymers; D012333:RNA, Messenger; D011094:Polyethyleneimine; D004247:DNA
keywords: cationic lipids; cationic polymers; cytosolic delivery; endosome escape; gene transfer; mRNA transfer