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Received December 6, 2004
Accepted January 17, 2005
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저분자량 키토산/유전자 나노콤플렉스 제조 및 이를 이용한 293 세포로의 전달
DNA Condensation and Delivery in 293 Cells Using Low Molecular Weight Chitosan/gene Nano-complex
중소기업진흥공단 중소기업연수원 생산기술연수실, 425-781 경기도 안산시 원곡동 931 1연세대학교 의과대학 연세심혈관연구소, 120-749 서울시 서대문구 신촌동 134 2연세대학교 화학공학과, 120-749 서울시 서대문구 신촌동 134
Office of Technical Instructors, Small Business Training Institute, 931, Wongok-dong, Ansan, Kyonggi-do 425-781, Korea 1Yonsei Cardiovascular Research Institute, College of Medicine, Yonsei University, 134, Shinchon-dong, Seodaemun-gu, Seoul 120-749, Korea 2Department of Chemical Engineering, Yonsei University, 134, Shinchon-dong, Seodaemun-gu, Seoul 120-749, Korea
Korean Chemical Engineering Research, April 2005, 43(2), 313-317(5), NONE Epub 9 May 2005
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Abstract
양이온성 고분자와 같은 합성 유전자 전달체들은 음이온성을 지닌 plasmid DNA와 쉽게 콤플렉스를 형성하는 경향이 있다. 이에 키토산은 유전자 전달체 시스템으로써 이용되어 질 수 있는 무한한 가능성을 지닌 polysaccharide이다. 저분자량 키토산이 DNA와 결합을 할 수 있는지 확인하기 위하여 전기영동장치를 이용하여 분석하였다. DLS(dynamic laser scattering)와 SEM(scanning electron microscopy)을 이용하여 키토산/DNA 콤플렉스의 크기와 모폴로지를 조사 하였다. 또한, 키토산의 분자량과 전하밀도가 콤플렉스의 크기와 결합된 DNA의 양에 어떻게 영향을 주는지 연구를 수행하였다. 저분자량 키토산은 실험과정에서 사용되는 양을 늘려갈수록 84-108%의 세포 생존율을 보임에 따라 그 독성이 무시할 정도가 됨을 확인할 수 있었다. 키토산/DNA 콤플렉스를 이용한 유전자 발현 효율 실험에서는 lipofecamine에 비해서는 낮은 값을 보였지만, naked DNA를 이용한 경우보다는 상대적으로 높은 값을 나타내었다. 키토산의 분자량에 따른 유전자 발현 효율 연구에서는 평균 분자량이 8,517인 키토산을 사용한 경우가 4,078의 분자 량을 이용한 실험 결과보다 높은 값을 보였고, 이는 키토산의 전하밀도가 유전자 발현 효율에 영향을 준다는 것을 확인할 수 있었다.
Synthetic gene carriers such as poly-cationic polymers easily form complexes with plasmid DNA which contains negative charge. Chitosan is a polysaccharide that demonstrates much potential as a gene delivery system. The ability of depolymerized chitosan to condense DNA was determined using electrophoresis. Dynamic laser scattering and scanning electron microscopy were used to examine the size and the morphology of the chitosan/DNA complex. Parameters such as chitosan molecular weight and charge density influenced the complex size and the DNA amount condensed with chitosan. The cell viabilities in the presence of chitosan ranged between 84-108% of the control in all experiments. Gene expression efficacy using chitosan/DNA complex was enhanced in 293 cells relative to that using naked DNA, although it was lower than that using lipofecamine. Transfection efficacy using low molecular weight chitosan (Mw=8,517) was higher than those of the control and the other chitosan (MW=4,078). The low molecular weight chitosan (MW=8,517) with a high charge density (18.32 mV) fulfilled the requirements for a suitable model gene delivery system with respect to the condensing ability of DNA, complex formation, and transfection efficacy.
References
Crystal RG, Nat. Med., 1(1), 15 (1995)
Pouton CW, Seymour LW, Adv. Drug Deliv. Rev., 34(1), 3 (1998)
Jones NA, Hill IRC, Stolnik S, Bignotti F, Davis SS, Garnet MC, Biochim Biophys Acta, 1517(1), 1 (2000)
Choi YH, Liu F, Park JS, Kim SW, Bioconjugate Chem., 9(6), 708 (1998)
Gebhart CL, Kabanov AV, J. Control. Release, 73(3), 401 (2001)
Haensler J, Szoka FC, Bioconjugate Chem., 4(5), 372 (1993)
Lim YB, Han SO, Kong HU, Pharm. Res., 17(7), 811 (2000)
Fiona C, MacLaughlin FC, Mumper RJ, Wang J, Tagliaferri JM, Gill I, Hinchcliffe M, Rolland AP, J. Control. Release, 56(3), 259 (1998)
Roy K, Mao HQ, Huang SK, Leong KW, Nat. Med., 5(4), 387 (1999)
Lee KY, Kwon IC, Kim YH, Jo WH, Jeon SY, J. Control. Release, 12(2), 213 (1998)
Mao HQ, Krishnendu R, Troung-Le VL, Kevin AJ, Kevin YL, Yan WJ, Thomas JA, Kam WL, J. Control. Release, 70(3), 399 (2001)
Peniston QP, Johnson EL, USP, No. 3,922,260, Nov., 25 (1975)
Pouton CW, Seymour LW, Adv. Drug Deliv. Rev., 34(1), 3 (1998)
Jones NA, Hill IRC, Stolnik S, Bignotti F, Davis SS, Garnet MC, Biochim Biophys Acta, 1517(1), 1 (2000)
Choi YH, Liu F, Park JS, Kim SW, Bioconjugate Chem., 9(6), 708 (1998)
Gebhart CL, Kabanov AV, J. Control. Release, 73(3), 401 (2001)
Haensler J, Szoka FC, Bioconjugate Chem., 4(5), 372 (1993)
Lim YB, Han SO, Kong HU, Pharm. Res., 17(7), 811 (2000)
Fiona C, MacLaughlin FC, Mumper RJ, Wang J, Tagliaferri JM, Gill I, Hinchcliffe M, Rolland AP, J. Control. Release, 56(3), 259 (1998)
Roy K, Mao HQ, Huang SK, Leong KW, Nat. Med., 5(4), 387 (1999)
Lee KY, Kwon IC, Kim YH, Jo WH, Jeon SY, J. Control. Release, 12(2), 213 (1998)
Mao HQ, Krishnendu R, Troung-Le VL, Kevin AJ, Kevin YL, Yan WJ, Thomas JA, Kam WL, J. Control. Release, 70(3), 399 (2001)
Peniston QP, Johnson EL, USP, No. 3,922,260, Nov., 25 (1975)
