DNA Targeting Sequence Improves Magnetic Nanoparticle-Based Plasmid DNA Transfection Efficiency in Model Neurons.
Efficient non-viral plasmid DNA transfection of most stem cells, progenitor cells and primary cell lines currently presents an obstacle for many applications within gene therapy research. From a standpoint of efficiency and cell viability, magnetic nanoparticle-based DNA transfection is a promising gene vectoring technique because it has demonstrated rapid and improved transfection outcomes when compared to alternative non-viral methods. Recently, our research group introduced oscillating magnet arrays that resulted in further improvements to this novel plasmid DNA (pDNA) vectoring technology. Continued improvements to nanomagnetic transfection techniques have focused primarily on magnetic nanoparticle (MNP) functionalization and transfection parameter optimization: cell confluence, growth media, serum starvation, magnet oscillation parameters, etc. Noting that none of these parameters can assist in the nuclear translocation of delivered pDNA following MNP-pDNA complex dissociation in the cell's cytoplasm, inclusion of a cassette feature for pDNA nuclear translocation is theoretically justified. In this study incorporation of a DNA targeting sequence (DTS) feature in the transfecting plasmid improved transfection efficiency in model neurons, presumably from increased nuclear translocation. This observation became most apparent when comparing the response of the dividing SH-SY5Y precursor cell to the non-dividing and differentiated SH-SY5Y neuroblastoma cells.
Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, FL 32611, USA. vernonmm@gmail.com.
Matthew M
MM
Vernon
School of Medicine and Dentistry, University of Rochester Medical Center, Rochester, NY 14642, USA. david_dean@urmc.rochester.edu.
David A
DA
Dean
Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, FL 32611, USA. jdobson@ufl.edu.|Department of Materials Science and Engineering, University of Florida, Gainesville, FL 32603, USA. jdobson@ufl.edu.|Institute for Cell Engineering & Regenerative Medicine (ICERM), University of Florida, Gainesville, FL 32611, USA. jdobson@ufl.edu.
Jon
J
Dobson
D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D006801:Humans; D008280:Magnetics / Q000379:methods; D058185:Magnetite Nanoparticles / Q000737:chemistry; D009474:Neurons / Q000378:metabolism*; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D014162:Transfection / Q000379:methods*
D058185:Magnetite Nanoparticles; D004247:DNA
DNA targeting sequence; SH-SY5Y cells; gene transfection; magnetic nanoparticles; neuroblastoma; nuclear localization; plasmid DNA; primary neurons