Development Of A Non-viral Gene Delivery Vector Based On The Dynein Light Chain Rp3 And The Tat Peptide.

dc.creatorFavaro, M T P
dc.creatorde Toledo, M A S
dc.creatorAlves, R F
dc.creatorSantos, C A
dc.creatorBeloti, L L
dc.creatorJanissen, R
dc.creatorde la Torre, L G
dc.creatorSouza, A P
dc.creatorAzzoni, A R
dc.date2014-Mar
dc.date2015-11-27T13:41:46Z
dc.date2015-11-27T13:41:46Z
dc.date.accessioned2018-03-29T01:19:33Z
dc.date.available2018-03-29T01:19:33Z
dc.identifierJournal Of Biotechnology. v. 173, p. 10-8, 2014-Mar.
dc.identifier1873-4863
dc.identifier10.1016/j.jbiotec.2014.01.001
dc.identifierhttp://www.ncbi.nlm.nih.gov/pubmed/24417903
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/201088
dc.identifier24417903
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1301321
dc.descriptionGene therapy and DNA vaccination trials are limited by the lack of gene delivery vectors that combine efficiency and safety. Hence, the development of modular recombinant proteins able to mimic mechanisms used by viruses for intracellular trafficking and nuclear delivery is an important strategy. We designed a modular protein (named T-Rp3) composed of the recombinant human dynein light chain Rp3 fused to an N-terminal DNA-binding domain and a C-terminal membrane active peptide, TAT. The T-Rp3 protein was successfully expressed in Escherichia coli and interacted with the dynein intermediate chain in vitro. It was also proven to efficiently interact and condense plasmid DNA, forming a stable, small (∼100nm) and positively charged (+28.6mV) complex. Transfection of HeLa cells using T-Rp3 revealed that the vector is highly dependent on microtubule polarization, being 400 times more efficient than protamine, and only 13 times less efficient than Lipofectamine 2000™, but with a lower cytotoxicity. Confocal laser scanning microcopy studies revealed perinuclear accumulation of the vector, most likely as a result of transport via microtubules. This study contributes to the development of more efficient and less cytotoxic proteins for non-viral gene delivery.
dc.description173
dc.description10-8
dc.languageeng
dc.relationJournal Of Biotechnology
dc.relationJ. Biotechnol.
dc.rightsfechado
dc.rightsCopyright © 2014 Elsevier B.V. All rights reserved.
dc.sourcePubMed
dc.subjectCytoplasmic Dyneins
dc.subjectGene Products, Tat
dc.subjectGene Transfer Techniques
dc.subjectGenetic Vectors
dc.subjectHela Cells
dc.subjectHumans
dc.subjectLipids
dc.subjectMicroscopy, Confocal
dc.subjectMicrotubules
dc.subjectMolecular Mimicry
dc.subjectPeptides
dc.subjectProtamines
dc.subjectRecombinant Proteins
dc.subjectTransfection
dc.subjectDynein Light Chain Rp3
dc.subjectGene Delivery
dc.subjectNon-viral Protein Vectors
dc.subjectTat
dc.titleDevelopment Of A Non-viral Gene Delivery Vector Based On The Dynein Light Chain Rp3 And The Tat Peptide.
dc.typeArtículos de revistas


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