Enhancement of efficiency of chitosan-based complexes for gene transfection with poly(γ-glutamic acid) by augmenting their cellular uptake and intracellular unpackage.
As a cationic polysaccharide, chitosan (CS) has been identified for its potential use as a non-viral vector for exogenous gene transfection. However, owing to their electrostatic interactions, CS complexes may cause difficulties in gene release upon their arrival at the site of action, thus limiting their transfection efficiency. In this work, an attempt is made to facilitate the release of a gene by incorporating a negatively-charged poly(γ-glutamic acid) (γPGA) into CS complexes in order to diminish their attractive interactions. The mechanisms of exploiting γPGA to enhance the transfection efficiency of CS complexes are elucidated. The feasibility of using this CS/γPGA-based system for DNA or siRNA transfer is explored as well. Additionally, potential of the CS/γPGA formulation to deliver disulfide bond-conjugated dual PEGylated siRNAs for multiple gene silencing is also examined. Moreover, the genetic use of pKillerRed-mem, delivered using complexes of CS and γPGA, to express a membrane-targeted KillerRed as an intrinsically generated photosensitizer for photodynamic therapy is described.
Department of Chemical Engineering and Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan, ROC.
Zi-Xian
ZX
Liao
Department of Biological Science and Technology and Department of Medical Research, China Medical University, Taichung, Taiwan, ROC.
Shu-Fen
SF
Peng
Department of Chemical Engineering and Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan, ROC.
Ya-Ling
YL
Chiu
Department of Chemical Engineering and Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan, ROC.
Chun-Wen
CW
Hsiao
Department of Chemical Engineering and Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan, ROC.
Hung-Yi
HY
Liu
Department of Chemical Engineering and Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan, ROC.
Woon-Hui
WH
Lim
Department of Chemical Engineering and Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan, ROC.
Hsiang-Ming
HM
Lu
Department of Chemical Engineering and Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan, ROC. Electronic address: hwsung@che.nthu.edu.tw.
Hsing-Wen
HW
Sung
D045744:Cell Line, Tumor; D002470:Cell Survival / Q000235:genetics; D048271:Chitosan / Q000737:chemistry* / Q000494:pharmacology; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D004337:Drug Carriers / Q000737:chemistry* / Q000494:pharmacology; D004355:Drug Stability; D004705:Endocytosis / Q000187:drug effects; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics; D056004:Molecular Dynamics Simulation; D011099:Polyglutamic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D014162:Transfection
D004337:Drug Carriers; D034741:RNA, Small Interfering; C485184:enhanced green fluorescent protein; C508771:killer red protein, Anthomedusae; C511775:poly(gamma-glutamic acid); D049452:Green Fluorescent Proteins; D011099:Polyglutamic Acid; D004247:DNA; D048271:Chitosan; D008156:Luciferases
Gene silencing; Gene therapy; Photodynamic therapy; poly(γ-glutamic acid); γ-glutamyl transpeptidase