Gene transfer using self-assembled ternary complexes of cationic magnetic nanoparticles, plasmid DNA and cell-penetrating Tat peptide.
Nonviral magnetofection facilitates gene transfer by using a magnetic field to concentrate magnetic nanoparticle-associated plasmid delivery vectors onto target cells. In light of the well-established effects of the Tat peptide, a cationic cell-penetrating peptide, that enhances the cytoplasmic delivery of a variety of cargos, we tested whether the combined use of magnetofection and Tat-mediated intracellular delivery would improve transfection efficiency. Through electrostatic interaction, gene transfer complexes were generated by mixing polyethylenimine-coated cationic magnetic iron beads with plasmid DNA, followed by addition of a bis(cysteinyl) histidine-rich Tat peptide. These ternary magnetofection complexes provided a 4-fold improvement in transgene expression at a dose of 1 microg of plasmid DNA per 20,000 cells over the binary complexes without the Tat peptide and transfected up to 60% of cells in vitro. The enhanced transfection efficiency was also observed in vivo in the rat spinal cord after lumbar intrathecal injection. Moreover, the injected ternary magnetofection complexes in the cerebrospinal fluid responded to a moving magnetic filed by shifting away from the injection site and mediating transgene expression in a remote region. Thus, our approach could potentially be useful for effective gene therapy treatments of localized diseases.
Song
Hai Peng
HP
Department of Biological Sciences, National University of Singapore, Singapore.
Yang
Jing Ye
JY
Lo
Seong Loong
SL
Wang
Yi
Y
Fan
Wei Min
WM
Tang
Xiao Sheng
XS
Xue
Jun Min
JM
Wang
Shu
S
D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival; D018014:Gene Transfer Techniques; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008280:Magnetics; D008297:Male; D053768:Metal Nanoparticles / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar; D013116:Spinal Cord / Q000378:metabolism; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000737:chemistry*
D010446:Peptide Fragments; D054322:tat Gene Products, Human Immunodeficiency Virus; D008156:Luciferases