Alkyl sulfonyl derivatized PAMAM-G2 dendrimers as nonviral gene delivery vectors with improved transfection efficiencies.
Amphiphilic dendrimer-based gene delivery vectors bearing peripheral alkyl sulfonyl hydrophobic tails were constructed using low-generation PAMAM-G2 as the core and functionalized by means of the aza-Michael type addition of its primary amino groups to vinylsulfone derivatives as an efficient tool for surface engineering. While the unmodified PAMAM-G2 was unable to efficiently transfect eukaryotic cells, functionalized PAMAM-G2 dendrimers were able to bind DNA at low N/P ratios, protect DNA from digestion with DNase I and showed high transfection efficiencies and low cytotoxicity. Dendrimers with a C18 alkyl chain produced transfection efficiencies up to 3.1 fold higher than LipofectAMINE™ 2000 in CHO-k1 cells. The dendriplexes based in functionalized PAMAM-G2 also showed the ability to retain their transfection properties in the presence of serum and the ability to transfect different eukaryotic cell lines such as Neuro-2A and RAW 264.7. Taking advantage of the vinylsulfone chemistry, fluorescent PAMAM-G2 derivatives of these vectors were prepared as molecular probes to determine cellular uptake and internalization through a clathrin-independent mechanism.
Departamento de Q. Orgánica, Facultad de Ciencia, Instituto de Biotecnología, Universidad de Granada, Granada, 18071, Spain.
Julia
J
Morales-Sanfrutos
Alicia
A
Megia-Fernandez
Fernando
F
Hernandez-Mateo
Ma Dolores
MD
Giron-Gonzalez
Rafael
R
Salto-Gonzalez
Francisco
F
Santoyo-Gonzalez
D000478:Alkylation; D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D050091:Dendrimers / Q000737:chemistry* / Q000494:pharmacology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000737:chemistry*; D051379:Mice; D015394:Molecular Structure; D013450:Sulfones / Q000737:chemistry*
D050091:Dendrimers; D013450:Sulfones