Investigation of aminoethyl methacrylate polymers for in vivo delivery of mRNA
Başlık:
Investigation of aminoethyl methacrylate polymers for in vivo delivery of mRNA
Yazar:
Esmer, Ayça, author.
Yazar Ek Girişi:
Fiziksel Tanımlama:
xii, 54 leaves: illustrarions, charts; 29 cm + 1 computer laser optical disc.
Özet:
There is a tremendous need for non-viral vectors for safe and efficient gene therapies. Especially the use of polymeric vectors for messenger RNA (mRNA) based gene therapies is limited. Within the scope of this thesis, it is aimed to perform a preliminary investigation on the in vivo transfection ability of a newly developed polymeric vector using zebrafish embryos in comparison with well-known lipidic and polymeric vectors. In line with this goal, the mRNA transfection ability of the block copolymer, poly(oligo(ethylene glycol) methacrylate)-b-poly(2- (amino)ethyl)amino)ethyl methacrylate, P(OEGMA)42-b-P(AEAEMA)48 along with Lipofectamine 3000 and branched polyethylene imine (PEI) (25 kDa) was examined in vivo on zebrafish embryos. A number of optimization experiments were first performed using naked mRNA or Lipofectamine-mRNA complexes to determine the best administration site and method, mRNA dose, type and development stage of embryos within the studied range. Considering the results obtained from optimization experiments, polyplexes formed with GFP-mRNA (2000 ng) and P(OEGMA)42-b-P(AEAEMA)48 at an N/P ratio of 3.6 or 7.3 were injected into the pericardial cavity of developing zebrafish embryos at 48 hours post fertilization to observe GFP expression. Naked mRNA, naked embryos, Lipofectamine-mRNA complex, and PEI-mRNA polyplexes were used for comparison. Samples were visualized 24 hours after injection using confocal microscopy and analyzed with Image J. The block copolymer showed transfection efficiency comparable with the golden standard polymeric vector PEI. The preliminary results obtained in this study pave the way for further investigations on the in vivo applications of P(OEGMA)42-b-P(AEAEMA)48 as a potential polymeric vector for mRNA-based gene therapies
Yazar Ek Girişi:
Tüzel Kişi Ek Girişi:
Tek Biçim Eser Adı:
Thesis (Masterl)-- İzmir Institute of Technology: Bioengineering.
İzmir Institute of Technology: Bioengineering. (Master).
Elektronik Erişim:
Access to Electronic Versiyon.