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Article: Revisit complexation between DNA and polyethylenimine - Effect of uncomplexed chains free in the solution mixture on gene transfection

TitleRevisit complexation between DNA and polyethylenimine - Effect of uncomplexed chains free in the solution mixture on gene transfection
Authors
KeywordsCellular internalization
DNA condensation
Free polyethylenimine
Gene delivery
Laser light scattering
Issue Date2011
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jconrel
Citation
Journal Of Controlled Release, 2011, v. 155 n. 1, p. 67-76 How to Cite?
Abstract
Our revisit of the complexation between anionic DNA and cationic polyethylenimine (PEI) in both water and phosphate buffered saline (PBS) by using a combination of laser light scattering (LLS) and gel electrophoresis confirms that nearly all the DNA chains are complexed with PEI to form polyplexes when the molar ratio of nitrogen from PEI to phosphate from DNA (N:P) reaches ~ 3, but the PEI/DNA polyplexes have a high in-vitro gene transfection efficiency only when N:P ≥ 10. Putting these two facts together, we not only conclude that this extra 7 portions of PEI chains are free in the solution mixture, but also confirmed that it is these free PEI chains that substantially promote the gene transfection no matter whether they are applied hours before or after the administration of the much less effective PEI/DNA polyplexes (N:P = 3). The uptake kinetics measured by flow cytometry shows that the addition of free PEI leads to a faster and more efficient cellular internalization of polyplexes, but these free PEI chains mainly contribute to the subsequent intracellular trafficking. In contrast, the bound PEI chains mainly play a role in the DNA condensation and protection, leading to a different thinking in the development of non-viral vectors. © 2010 Elsevier B.V. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/129117
ISSN
2013 Impact Factor: 7.261
2013 SCImago Journal Rankings: 2.494
ISI Accession Number ID
Funding AgencyGrant Number
National Natural Scientific Foundation of China (NNSFC)50773077
20934005
Hong Kong Special Administration Region Earmarked (RGC)CUHK4037/07P
2160331
CUHK4046/08P
2160365
CUHK4039/08P
2160361
CUHK4042/09P
2160396
Funding Information:

The financial support of the National Natural Scientific Foundation of China (NNSFC) Projects (50773077 and 20934005) and the Hong Kong Special Administration Region Earmarked (RGC) Projects (CUHK4037/07P, 2160331; CUHK4046/08P, 2160365; CUHK4039/08P, 2160361; and CUHK4042/09P, 2160396) is gratefully acknowledged.

References

 

Author Affiliations
  1. The University of Hong Kong
  2. University of Science and Technology of China
  3. Chinese University of Hong Kong
DC FieldValueLanguage
dc.contributor.authorYue, Yen_HK
dc.contributor.authorJin, Fen_HK
dc.contributor.authorDeng, Ren_HK
dc.contributor.authorCai, Jen_HK
dc.contributor.authorChen, Yen_HK
dc.contributor.authorLin, MCMen_HK
dc.contributor.authorKung, HFen_HK
dc.contributor.authorWu, Cen_HK
dc.date.accessioned2010-12-23T08:32:44Z-
dc.date.available2010-12-23T08:32:44Z-
dc.date.issued2011en_HK
dc.identifier.citationJournal Of Controlled Release, 2011, v. 155 n. 1, p. 67-76en_HK
dc.identifier.issn0168-3659en_HK
dc.identifier.urihttp://hdl.handle.net/10722/129117-
dc.description.abstractOur revisit of the complexation between anionic DNA and cationic polyethylenimine (PEI) in both water and phosphate buffered saline (PBS) by using a combination of laser light scattering (LLS) and gel electrophoresis confirms that nearly all the DNA chains are complexed with PEI to form polyplexes when the molar ratio of nitrogen from PEI to phosphate from DNA (N:P) reaches ~ 3, but the PEI/DNA polyplexes have a high in-vitro gene transfection efficiency only when N:P ≥ 10. Putting these two facts together, we not only conclude that this extra 7 portions of PEI chains are free in the solution mixture, but also confirmed that it is these free PEI chains that substantially promote the gene transfection no matter whether they are applied hours before or after the administration of the much less effective PEI/DNA polyplexes (N:P = 3). The uptake kinetics measured by flow cytometry shows that the addition of free PEI leads to a faster and more efficient cellular internalization of polyplexes, but these free PEI chains mainly contribute to the subsequent intracellular trafficking. In contrast, the bound PEI chains mainly play a role in the DNA condensation and protection, leading to a different thinking in the development of non-viral vectors. © 2010 Elsevier B.V. All rights reserved.en_HK
dc.languageengen_US
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jconrelen_HK
dc.relation.ispartofJournal of Controlled Releaseen_HK
dc.subjectCellular internalizationen_HK
dc.subjectDNA condensationen_HK
dc.subjectFree polyethylenimineen_HK
dc.subjectGene deliveryen_HK
dc.subjectLaser light scatteringen_HK
dc.titleRevisit complexation between DNA and polyethylenimine - Effect of uncomplexed chains free in the solution mixture on gene transfectionen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0168-3659&volume=155&issue=1&spage=67&epage=76&date=2011&atitle=Revisit+complexation+between+DNA+and+polyethylenimine:+effect+of+uncomplexed+chains+free+in+the+solution+mixture+on+gene+transfection-
dc.identifier.emailLin, MCM:mcllin@hkucc.hku.hken_HK
dc.identifier.authorityLin, MCM=rp00746en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.jconrel.2010.10.028en_HK
dc.identifier.pmid21056067en_HK
dc.identifier.scopuseid_2-s2.0-79953839673en_HK
dc.identifier.hkuros183297en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-79953839673&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume155en_HK
dc.identifier.issue1en_HK
dc.identifier.spage67en_HK
dc.identifier.epage76en_HK
dc.identifier.isiWOS:000295998200012-
dc.publisher.placeNetherlandsen_HK
dc.identifier.scopusauthoridYue, Y=35088987100en_HK
dc.identifier.scopusauthoridJin, F=35315971100en_HK
dc.identifier.scopusauthoridDeng, R=24467381600en_HK
dc.identifier.scopusauthoridCai, J=36668119900en_HK
dc.identifier.scopusauthoridChen, Y=24075600300en_HK
dc.identifier.scopusauthoridLin, MCM=7404816359en_HK
dc.identifier.scopusauthoridKung, HF=7402514190en_HK
dc.identifier.scopusauthoridWu, C=35075235200en_HK
dc.identifier.citeulike8215993-

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