title: Targeted delivery of in situ PCR-amplified Sleeping Beauty transposon genes to cancer cells with lipid-based nanoparticle-like protocells.
abstract: A Sleeping Beauty (SB) transposon system is made of a transposon plasmid (containing gene encoding a desired functional or therapeutic protein) and a transposase plasmid (encoding an enzyme capable of cutting and pasting the gene into the host cell genome). It is a kind of natural, nonviral gene delivery vehicle, which can achieve efficient genomic insertion, providing long-term transgenic expression. However, before the SB transposon system could play a role in promoting gene expression, it has to be delivered efficiently first across cell membrane and then into cell nuclei. Towards this end, we used a nanoparticle-like lipid-based protocell, a closed bilayer of the neutral lipids with the DNA encapsulated inside, to deliver the SB transposon system to cancer cells. The SB transposon system was amplified in situ inside the protocells by a polymerase chain reaction (PCR) process, realizing more efficient loading and delivery of the target gene. To reach a high transfection efficiency, we introduced two targeting moieties, folic acid (FA) as a cancer cell-targeting motif and Dexamethasone (DEX) as a nuclear localization signaling molecule, into the protocells. As a result, the FA enabled the modified targeting protocells to deliver the DNA into the cancer cells with an increased efficiency and the DEX promoted the DNA to translocate to cell nuclei, eventually leading to the increased chromosome insertion efficiency of the SB transposon. In vivo study strongly suggested that the transfection efficiency of FA-modified protocells in the tumor tissue was much higher than that in other tissues, which was consistent with the in vitro results. Our studies implied that with the targeting ligand modification, the protocells could be utilized as an efficient targeting gene carrier. Since the protocells were made of neutral lipids without cationic charges, the cytotoxicity of protocells was significantly lower than that of traditional cationic gene carriers such as cationic liposomes and polyethylenimine, enabling the protocells to be employed in a wider dosage range in gene therapy. Our work shows that the protocells are a promising gene carrier for future clinical applications.
authors.affiliation: School of Life Science and Medicine, Dalian University of Technology, Panjin, Liaoning 124221, China; Department of Chemistry and Biochemistry, Stephenson Life Sciences Research Center, University of Oklahoma, OK, 73019, USA. Electronic address: makunonline@163.com.
authors.forename: Kun
authors.initials: K
authors.lastname: Ma
authors.affiliation: School of Life Science and Medicine, Dalian University of Technology, Panjin, Liaoning 124221, China.
authors.forename: Duo
authors.initials: D
authors.lastname: Fu
authors.affiliation: School of Life Science and Medicine, Dalian University of Technology, Panjin, Liaoning 124221, China.
authors.forename: Dongli
authors.initials: D
authors.lastname: Yu
authors.affiliation: School of Life Science and Medicine, Dalian University of Technology, Panjin, Liaoning 124221, China.
authors.forename: Changhao
authors.initials: C
authors.lastname: Cui
authors.affiliation: School of Life Science and Medicine, Dalian University of Technology, Panjin, Liaoning 124221, China.
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.affiliation: School of Life Science and Medicine, Dalian University of Technology, Panjin, Liaoning 124221, China.
authors.forename: Zhaoming
authors.initials: Z
authors.lastname: Guo
authors.affiliation: Department of Chemistry and Biochemistry, Stephenson Life Sciences Research Center, University of Oklahoma, OK, 73019, USA; School of Materials Science and Engineering, Zhejiang University, Hangzhou, Zhejiang, 310027, China. Electronic address: cbmao@ou.edu.
authors.forename: Chuanbin
authors.initials: C
authors.lastname: Mao
mesh_terms: D000818:Animals; D058045:Artificial Cells / Q000737:chemistry*; D045744:Cell Line, Tumor; D004251:DNA Transposable Elements / Q000235:genetics; D005260:Female; D018390:Gene Targeting / Q000379:methods; D015316:Genetic Therapy / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D008055:Lipids / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D053769:Nanocapsules / Q000008:administration & dosage / Q000737:chemistry*; D009374:Neoplasms, Experimental / Q000235:genetics* / Q000628:therapy*; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D014162:Transfection / Q000379:methods; D019895:Transposases / Q000008:administration & dosage / Q000235:genetics*; D016896:Treatment Outcome
chemical_list: D004251:DNA Transposable Elements; D008055:Lipids; D053769:Nanocapsules; D019895:Transposases; C494471:sleeping beauty transposase, human
keywords: Cancer cells; Non-viral gene carrier; Protocells; Sleeping Beauty transposon; Targeted delivery