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Development of Non-Viral Vectors for Gene Therapy and DNA Vaccination: Overcoming Barriers to Gene Delivery

Grant number: 09/12635-5
Support type:Scholarships in Brazil - Master
Effective date (Start): August 01, 2010
Effective date (End): May 31, 2012
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal researcher:Adriano Rodrigues Azzoni
Grantee:Marianna Teixeira de Pinho Favaro
Home Institution: Centro de Biologia Molecular e Engenharia Genética (CBMEG). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil


A major limiting factor for the development of more efficient protocols of gene therapy and DNA vaccination arises from the low efficiency of gene transfer by non-viral-vectors. This comes mainly from the difficulty to transport foreign DNA from outside into the target cell's nucleus, due to the presence of several physical, enzymatic and diffusional barriers. The main objective of the project here proposed is the development of multifunctional non-viral vectors ("artificial virus") capable of efficiently delivering genetic material. By introducing new domains to the fusion proteins currently under study by our group (TcTex-1 e RP3), we expect to evaluate systematically these proteins´ ability to help overcome the existing barriers. The increased efficiency of entry into the cell and translocation to the nucleus will be studied through the inclusion of "membrane-active" domains (Tat and Penetratine). The escape of vectors retained in endosomes will be evaluated through the use of histidine sequences. Intracellular transport of pDNA through microtubules will be studied by exploring the natural capacity of the Dynein motor protein to transport cargo from the periphery to the interior of cells. At all stages of the study, the number of pDNA vectors present in the interior and nucleus of transfected cells will be quantified by Real Time PCR, and transfection efficiency will be measured via the expression of the reporter genes Luciferase and GFP. As a result, it is expected to raise important information about some of the main mechanisms involved in crucial steps of intracellular trafficking of transgenes. Other expected result is the development of multifunctional vectors and reduce the difference of efficiency that currently exist between viral and non-viral vectors.

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Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
FAVARO, Marianna Teixeira de Pinho. Development of non viral vectors for gene delivery based on dynein light chain Rp3. 2012. Master's Dissertation - Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia Campinas, SP.

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