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Article: Effective endogenous gene silencing mediated by pH responsive peptides proceeds via multiple pathways

TitleEffective endogenous gene silencing mediated by pH responsive peptides proceeds via multiple pathways
Authors
KeywordsEndocytosis
GAPDH
Gene silencing
pH responsive peptides
siRNA
Issue Date2012
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jconrel
Citation
Journal Of Controlled Release, 2012, v. 158 n. 2, p. 293-303 How to Cite?
AbstractCationic amphipathic histidine rich peptides possess high plasmid DNA and siRNA delivery capabilities. To further understand the pH responsive siRNA delivery process and evaluate the capabilities of such peptides we have investigated their ability to mediate specific silencing of endogenous GAPDH gene activity in MCF-7 and A549 cells and compared this with plasmid DNA delivery. A substantial and selective reduction of both GAPDH activity and expression was achieved using pH responsive peptide vectors, which compared favourably with that mediated by commercially available non-viral vectors in terms of efficacy and toxicity. Furthermore, by comparing the efficacy of both gene delivery and silencing mediated by a series of such peptides, their sensitivities to known inhibitors of endocytotic processes, and their route of uptake via confocal live cell imaging, we show that both plasmid DNA and siRNA are internalised via endocytosis. However siRNA entry facilitated by LAH4-L1, proceeds via a cholesterol dependent mechanism, in contrast to DNA transfer which is associated with clathrin dependent endocytosis. Furthermore, using peptides that respond at increasingly acidic pH, we demonstrate that the route of entry for the siRNA that ultimately mediates silencing is peptide specific and whilst some pH responsive peptides promote the escape of labelled siRNA from endosomes, others may promote entry via alternative mechanisms. © 2011 Elsevier B.V. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/146797
ISSN
2015 Impact Factor: 7.441
2015 SCImago Journal Rankings: 2.827
PubMed Central ID
ISI Accession Number ID
Funding AgencyGrant Number
Medical Research CouncilNIRG G0801072/87482
Wellcome Trust
University of Hong Kong
Funding Information:

This work was supported by the Medical Research Council (NIRG G0801072/87482 to AJM), the Wellcome Trust (VIP Award to AJM) and the University of Hong Kong Seed Funding Programme (JKWL). JKWL was the holder of a Maplethorpe Postdoctoral Fellowship of the University of London. JKWL and AJM thank Ms Mia So (Department of Pharmacology and Pharmacy, The University of Hong Kong) for her help in DNA transfection study, Mr Tony Chan (Department of Anatomy, The University of Hong Kong) and Ms Jing Guo (Faculty Core Facility, LKS Faculty of Medicine, The University of Hong Kong) for their assistance in the live cell confocal imaging experiment. We are grateful to Dr Antoine Kichler for critical reading of the manuscript.

References

 

DC FieldValueLanguage
dc.contributor.authorLam, JKWen_HK
dc.contributor.authorLiang, Wen_HK
dc.contributor.authorLan, Yen_HK
dc.contributor.authorChaudhuri, Pen_HK
dc.contributor.authorChow, MYTen_HK
dc.contributor.authorWitt, Ken_HK
dc.contributor.authorKudsiova, Len_HK
dc.contributor.authorMason, AJen_HK
dc.date.accessioned2012-05-15T02:01:49Z-
dc.date.available2012-05-15T02:01:49Z-
dc.date.issued2012en_HK
dc.identifier.citationJournal Of Controlled Release, 2012, v. 158 n. 2, p. 293-303en_HK
dc.identifier.issn0168-3659en_HK
dc.identifier.urihttp://hdl.handle.net/10722/146797-
dc.description.abstractCationic amphipathic histidine rich peptides possess high plasmid DNA and siRNA delivery capabilities. To further understand the pH responsive siRNA delivery process and evaluate the capabilities of such peptides we have investigated their ability to mediate specific silencing of endogenous GAPDH gene activity in MCF-7 and A549 cells and compared this with plasmid DNA delivery. A substantial and selective reduction of both GAPDH activity and expression was achieved using pH responsive peptide vectors, which compared favourably with that mediated by commercially available non-viral vectors in terms of efficacy and toxicity. Furthermore, by comparing the efficacy of both gene delivery and silencing mediated by a series of such peptides, their sensitivities to known inhibitors of endocytotic processes, and their route of uptake via confocal live cell imaging, we show that both plasmid DNA and siRNA are internalised via endocytosis. However siRNA entry facilitated by LAH4-L1, proceeds via a cholesterol dependent mechanism, in contrast to DNA transfer which is associated with clathrin dependent endocytosis. Furthermore, using peptides that respond at increasingly acidic pH, we demonstrate that the route of entry for the siRNA that ultimately mediates silencing is peptide specific and whilst some pH responsive peptides promote the escape of labelled siRNA from endosomes, others may promote entry via alternative mechanisms. © 2011 Elsevier B.V. All rights reserved.en_HK
dc.languageeng-
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.subjectEndocytosisen_HK
dc.subjectGAPDHen_HK
dc.subjectGene silencingen_HK
dc.subjectpH responsive peptidesen_HK
dc.subjectsiRNAen_HK
dc.titleEffective endogenous gene silencing mediated by pH responsive peptides proceeds via multiple pathwaysen_HK
dc.typeArticleen_HK
dc.identifier.emailLam, JKW: jkwlam@hku.hken_HK
dc.identifier.authorityLam, JKW=rp01346en_HK
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1016/j.jconrel.2011.11.024en_HK
dc.identifier.pmid22138072-
dc.identifier.pmcidPMC3309421-
dc.identifier.scopuseid_2-s2.0-84858701389en_HK
dc.identifier.hkuros199411-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-84858701389&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume158en_HK
dc.identifier.issue2en_HK
dc.identifier.spage293en_HK
dc.identifier.epage303en_HK
dc.identifier.isiWOS:000302607000014-
dc.publisher.placeNetherlandsen_HK
dc.identifier.scopusauthoridLam, JKW=8404243000en_HK
dc.identifier.scopusauthoridLiang, W=37017317100en_HK
dc.identifier.scopusauthoridLan, Y=36159603200en_HK
dc.identifier.scopusauthoridChaudhuri, P=54885464600en_HK
dc.identifier.scopusauthoridChow, MYT=54886510500en_HK
dc.identifier.scopusauthoridWitt, K=54885529300en_HK
dc.identifier.scopusauthoridKudsiova, L=21933984000en_HK
dc.identifier.scopusauthoridMason, AJ=8861864900en_HK
dc.identifier.citeulike10100448-

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