Gold nanoparticle-DNA conjugates for oligonucleotide vectorization towards gene silencing

Bibliographic Details
Main Author: Almeida, Carina Marisa dos Santos
Publication Date: 2011
Format: Master thesis
Language: eng
Source: Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
Download full: http://hdl.handle.net/10362/6212
Summary: Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina
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spelling Gold nanoparticle-DNA conjugates for oligonucleotide vectorization towards gene silencingGene silencingAntisense DNAGold nanoparticlesDNA triple helicesNanovectorizationDissertação para obtenção do Grau de Mestre em Genética Molecular e BiomedicinaThe main objective of the work presented in this thesis was to develop a gene silencing system by taking advantage of the nanovectorization capability and optical properties of gold nanoparticles. The idea is based on the construction of a DNA structure containing a therapeutic oligonucleotide with the ability to form Hoogsteen hydrogen bonds with double-stranded DNA, producing a DNA triple helix, besides silencing the gene of interest. Hoogsteen bonds, more unstable than the conventional Watson-Crick bonds, permit the achievement of lower melting temperatures. This attribute, coupled with the ability to generate heat by laser irradiation of the gold nanoparticles used, will allow the release of the therapeutic oligonucleotide and subsequent gene silencing without significant increase in the medium’s temperature. Thus, the thesis comprises three major sections: structure design and formation, vectorization, and gene expression silencing; the tasks involved in each of these sections were conducted in parallel. The design of the obtained structure took into account the desired melting temperature, stability at physiological conditions of the sequence-forming nucleotides, the number of Hoogsteen bonds and ionic conditions. To evaluate the formation of this structure, spectroscopic techniques were mainly used: FRET analysis and ultraviolet melting curves. Both approaches allowed the identification of interactions in the presence of therapeutic oligonucleotide compared with its absence, which may indicate structure formation. In addition, melting curves allowed the determination of the temperature of release of this oligonucleotide – 40ºC. The double-stranded DNA functionalization to gold nanoparticles has been achieved, but there was no difference in electrophoretic migration when the three oligonucleotides were present. However, the therapeutic oligonucleotide was able to efficiently inhibit gene expression in in vitro transcription and translation assays with efficiency up to 95% and 60% respectively.Faculdade de Ciências e TecnologiaBaptista, PedroRUNAlmeida, Carina Marisa dos Santos2011-11-07T11:43:06Z20112011-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/6212enginfo:eu-repo/semantics/openAccessreponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiainstacron:RCAAP2024-05-22T17:09:32Zoai:run.unl.pt:10362/6212Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireinfo@rcaap.ptopendoar:https://opendoar.ac.uk/repository/71602025-05-28T16:40:31.430413Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiafalse
dc.title.none.fl_str_mv Gold nanoparticle-DNA conjugates for oligonucleotide vectorization towards gene silencing
title Gold nanoparticle-DNA conjugates for oligonucleotide vectorization towards gene silencing
spellingShingle Gold nanoparticle-DNA conjugates for oligonucleotide vectorization towards gene silencing
Almeida, Carina Marisa dos Santos
Gene silencing
Antisense DNA
Gold nanoparticles
DNA triple helices
Nanovectorization
title_short Gold nanoparticle-DNA conjugates for oligonucleotide vectorization towards gene silencing
title_full Gold nanoparticle-DNA conjugates for oligonucleotide vectorization towards gene silencing
title_fullStr Gold nanoparticle-DNA conjugates for oligonucleotide vectorization towards gene silencing
title_full_unstemmed Gold nanoparticle-DNA conjugates for oligonucleotide vectorization towards gene silencing
title_sort Gold nanoparticle-DNA conjugates for oligonucleotide vectorization towards gene silencing
author Almeida, Carina Marisa dos Santos
author_facet Almeida, Carina Marisa dos Santos
author_role author
dc.contributor.none.fl_str_mv Baptista, Pedro
RUN
dc.contributor.author.fl_str_mv Almeida, Carina Marisa dos Santos
dc.subject.por.fl_str_mv Gene silencing
Antisense DNA
Gold nanoparticles
DNA triple helices
Nanovectorization
topic Gene silencing
Antisense DNA
Gold nanoparticles
DNA triple helices
Nanovectorization
description Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina
publishDate 2011
dc.date.none.fl_str_mv 2011-11-07T11:43:06Z
2011
2011-01-01T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/6212
url http://hdl.handle.net/10362/6212
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Faculdade de Ciências e Tecnologia
publisher.none.fl_str_mv Faculdade de Ciências e Tecnologia
dc.source.none.fl_str_mv reponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
instacron:RCAAP
instname_str FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
instacron_str RCAAP
institution RCAAP
reponame_str Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
collection Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
repository.name.fl_str_mv Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
repository.mail.fl_str_mv info@rcaap.pt
_version_ 1833596104166342656

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