Poly(amidoamine)s as potential nonviral vectors: ability to form interpolyelectrolyte complexes and to mediate transfection in vitro.
Poly(amidoamine)s (PAAs) are water-soluble polymers that display pH-dependent membrane activity. PAAs have the potential to act as a synthetic alternative to fusogenic peptides and thus promote endosomal escape. The purpose of this study was to investigate for the first time whether PAA have the ability to complex DNA, protect it from nuclease degradation and to promote transfection in vitro. PAAs ISA 1 (Mn 6900) and ISA 23 (Mn 10,500) and their 2-phenylethylamine containing analogues ISA 4 and ISA 22 (Mn approximately 8000) were studied. All PAAs retarded the electrophoretic mobility of lambda Hind III DNA demonstrating interpolyelectrolyte complex (IPEC) formation and toroids of 80-150 nm in diameter (10:1 polymer excess) were visible using TEM. DNase II inhibition was observed. At a polymer:DNA ratio of 10:1, this was ISA 1(89.6 +/- 6.1%), ISA 4 (92.2 +/- 11.2%), ISA 22 (69.4 +/- 3.7%), and ISA 23 (58.0 +/- 10.0%). PAAs demonstrated the ability to mediate pSV beta-galactosidase transfection of HepG2 cells. At a vector:DNA mass ratio of 5:1, ISA 23 showed equivalent transfection ability compared with polyethylenimine and LipofectIN and was more effective than LipofectACE. These properties suggest that PAAs warrant further development as endosomolytic vectors.
Richardson
S C
SC
Centre for Polymer Therapeutics, Welsh School of Pharmacy, Cardiff University, King Edward VII Avenue, Cardiff CF10 3XF, U.K.
Pattrick
N G
NG
Man
Y K
YK
Ferruti
P
P
Duncan
R
R
D002460:Cell Line; D004337:Drug Carriers / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D015394:Molecular Structure; D008970:Molecular Weight; D009757:Nylons / Q000737:chemistry*; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D011073:Polyamines / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry; D014162:Transfection
D004337:Drug Carriers; D046911:Macromolecular Substances; D009757:Nylons; D011073:Polyamines; D011108:Polymers