Application of Abf2 and MC1 proteins in preparation of short circular DNA for cell transfection

Abstract

Jednu od metoda molekulske terapije za modificiranje aktivnosti transkripcijskih faktora čine DNA-mamci (duljine manje od 100 pb). Do sada opisani oblici DNA-mamaca lako se degradiraju endonukleazama. Kružne molekule DNA su rezistentne na taj tip degradacije. Ipak, kratke molekule DNA (kraće od oko 150 pb) ne mogu se cirkularizirati samo uz upotrebu DNA-ligaze U ovom radu opisana je prva uspješna cirkularizacija dvolančane molekule DNA duljine 95 pb uz upotrebu proteina za savijanje DNA (Abf2 i MC1). Isto tako je konstruirana kružna DNA upotrijebljena za transfekciju stanica u kulturi.One of the molecular therapy methods for transcription factor activity modification are DNA-decoys (molecules shorter than 100 bp). All types of DNA-decoys that have been described so far are easily degraded by exonucleases. Circular DNA molecules are, however, resistant to exonuclease activity. Also, it is impossible to create circular DNA from linear dsDNA that is relatively shorter than 150 bp. This is the first published work with described circularization of 95 bp long dsDNA by using Abf2 protein and MC1 protein. The constructed circular DNA was used to transfect cells in cell culture