• Bulletin of the Korean Chemical Society
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• v.27 no.9
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• pp.1335-1340
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• 2006

It has been found that a number of diseases are associated with mutations in the mitochondrial DNA. Therapeutic gene delivery to mitochondria has been suggested as a clinical option for these diseases. In this study, we developed a gene carrier to mitochondria by the conjugation of mitochondrial leader peptide (LP) to polyethylenimine (PEI). Mitochondrial LP conjugated PEI (PEI-LP) was synthesized with low molecular weight PEI (2,000 Da, PEI2K). Gel retardation assay showed that PEI2K-LP formed complexes at a 1.0/1 weight ratio. In addition, PEI2K-LP protected DNA from the enzymatic degradation for at least 60 min, while naked DNA was completely degraded within 20 min. PEI2K-LP was compared with LP conjugated high molecular weight PEI (25,000 Da, PEI25K) in terms of toxicity and delivery efficiency. MTT assay showed that PEI2K-LP had much lower cytotoxicity than PEI25K-LP to 293 cells. In addition, cell-free DNA delivery assay showed that PEI2K-LP delivered more DNA to mitochondria at a 1.8/1 weight ratio than naked DNA or PEI. This result suggests that PEI2K-LP may be useful for the development of mitochondrial gene therapy system with lower cytotoxicity.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.4
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• pp.269-273
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• 2001

This study presents a new formulation method for improving DNA transfection effi-ciency using a fusogenic peptide and polyethylene glycol-grafted polyethylenimine. Succinimidyls succinate polyethylene glycol (PEG-SSA) was conjugated with polyethylenimine(PEL). PEL is well known for a good endosomal escaping and DNA condensign agent. The positively charged syn-thetic fusogenic peptide, KALA was coated on the negatively charged PEG-g-PEI/DNA and PEI/DNA complexes. The KALA/PEI/ DNA complexes exhibited aggregation behavior at higher KALA coating amount with an effective diameter of around 1,000 nm. However, the LALA/PEG-g-PEI/DNA complexes were 100-300 nm in size with a surface zeta-potential (ζ)value of about +20mV. The conjugated PEG molecules suppressed any KALA-mediated inter-particle aggregation, and thereby improved the transfection efficiency, Consequently, the transfection efficiency of the KALA/PEG-g-PEI/DNA complexes was obtained by utilizing both the fusogenic activity of KALA and the steric repulsion effect of PEC.

• Archives of Pharmacal Research
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• v.28 no.6
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• pp.722-729
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• 2005

With the aim to improve the specificity and to reduce the cytotoxicity of polyethylenimine (PEI), we have synthesized the conjugates of the branched PEI (25 kDa) with transferrin. The trans-ferrin-PEI (TP) conjugates with five compositions were synthesized using periodate oxidation method and confirmed by FT-IR spectroscopy and gel permeation chromatography. The free amine contents of TP conjugates, which were able to condense and deliver DNA, increased as the amount of PEI increased. TP/DNA polyplexes were characterized by measuring gel elec-trophoresis, ethidium bromide fluorescence quenching, particle size and zeta potential of complexes. Complete complexation of the polyplexes was observed above the N/P ratio of 5 in TP/DNA, and above 3 in PEI/DNA, respectively. The zeta potential of the complexes decreased as the amount of transferrin in TP conjugates increased. Transfection efficiency of TP conjugates was evaluated in HeLa cell and Jurkat cell systems. Among the five compositions of TP conjugates, TP-2 system mediated a higher $\beta$-galactosidase gene expression than PEI system in Jurkat cell which was known to express elevated numbers of transferrin receptors. From the results of the cell viability based on MTT assay, TP conjugates showed lower cytotoxicity com-pared with the PEI system. We expect that the TP conjugate can be used efficiently as a non-viral gene delivery vector.

• Journal of Pharmaceutical Investigation
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• v.35 no.1
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• pp.17-23
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• 2005

Polyethylenimine (PEI) has been used as cationic polymers for efficient gene transfer without the need for endosomolytic agents. Various kinds of PEIs with different molecular weight were tested in order to investigate the effects of the molecular weight of PEI on the transfection efficiency and cell cytotoxicity. The ${\beta}-galactosidase$ expression $(pCMV-{\beta}-gal)$ plasmid was used as a model DNA. Complex formation between PEI and pDNA was assessed by 1% agarose gel electrophoresis method. Particle size and zeta-potential of complexes were determined by electrophoretic light scattering spectrometer. In vitro transfection efficiency was assayed by measuring ${\beta}-galactosidase$ activity. Cell cytotoxicity was determined by MTT assay. Particle sizes of the complexes became smaller on increasing molecular weights of PEI and N/P ratios. Surface potential of complexes was increased as the molecular weight of PEI increased. Transfection efficiency of $pCMV-{\beta}-ga1$ on the HEK 293 cells was greatest with PEI 25 K system but having the lowest cell viability. PEI with high molecular weight showed higher transfection efficiency and cell viability than PEI with low molecular weight.