J F A Valente1,2, P Pereira1,3, A Sousa1, J A Queiroz1, F Sousa1. 1. CICS-UBI-Health Sciences Research Centre, Universidade da Beira Interior, Avenida Infante D. Henrique, 6200-506 Covilhã, Portugal. 2. CDRsp-Centre Rapid and Sustainable Product Development, Polytechnic Institute of Leiria, 2411-901 Leiria, Portugal. 3. CEMMPRE, Department of Chemical Engineering, University of Coimbra, Rua Sílvio Lima-Pólo II, 3030-790 Coimbra, Portugal.
Abstract
Gene therapy could be simply defined as a strategy for the introduction of a functional copy of desired genes in patients, to correct some specific mutation and potentially treat the respective disorder. However, this straightforward definition hides very complex processes related to the design and preparation of the therapeutic genes, as well as the development of suitable gene delivery systems. Within non-viral vectors, polymeric nanocarriers have offered an ideal platform to be applied as gene delivery systems. Concerning this, the main goal of the study was to do a systematic evaluation on the formulation of pDNA delivery systems based on the complexation of different sized plasmids with chitosan (CH) or polyethyleneimine (PEI) polymers to search for the best option regarding encapsulation efficiency, surface charge, size, and delivery ability. The cytotoxicity and the transfection efficiency of these systems were accessed and, for the best p53 encoding pDNA nanosystems, the ability to promote protein expression was also evaluated. Overall, it was showed that CH polyplexes are more efficient on transfection when compared with the PEI polyplexes, resulting in higher P53 protein expression. Cells transfected with CH/p53-pDNA polyplexes presented an increase of around 54.2% on P53 expression, while the transfection with the PEI/p53-pDNA polyplexes resulted in a 32% increase.
Gene therapy could be simply defined as a strategy for tpan> class="Chemical">he introduction of a functional copy of desired genes in patients, to correct some specific mutation and potentially treat the respective disorder. However, this straightforward definition hides very complex processes related to the design and preparation of the therapeutic genes, as well as the development of suitable gene delivery systems. Within non-viral vectors, polymeric nanocarriers have offered an ideal platform to be applied as gene delivery systems. Concerning this, the main goal of the study was to do a systematic evaluation on the formulation of pDNA delivery systems based on the complexation of different sized plasmids with chitosan (CH) or polyethyleneimine (PEI) polymers to search for the best option regarding encapsulation efficiency, surface charge, size, and delivery ability. Thecytotoxicity and the transfection efficiency of these systems were accessed and, for the best p53 encoding pDNA nanosystems, the ability to promote protein expression was also evaluated. Overall, it was showed that CH polyplexes are more efficient on transfection when compared with thePEI polyplexes, resulting in higher P53 protein expression. Cells transfected with CH/p53-pDNA polyplexes presented an increase of around 54.2% on P53 expression, while the transfection with thePEI/p53-pDNA polyplexes resulted in a 32% increase.
Authors: P Kreiss; B Cameron; R Rangara; P Mailhe; O Aguerre-Charriol; M Airiau; D Scherman; J Crouzet; B Pitard Journal: Nucleic Acids Res Date: 1999-10-01 Impact factor: 16.971
Authors: Feiyang Li; Jun Wu; Daiye Li; Liuzhi Hao; Yanqun Li; Dan Yi; Kelvin W K Yeung; Di Chen; William W Lu; Haobo Pan; Tak Man Wong; Xiaoli Zhao Journal: J Nanobiotechnology Date: 2022-03-15 Impact factor: 10.435