Non-viral gene delivery systems have great potential for safe and efficient gene therapy, while inefficient cellular and nuclear uptake remain as the major hurdles. Novel approaches are needed to enhance the transfection efficiency of non-viral vectors. In accordance with this need, the objective of this study was to construct a non-viral vector that could achieve gene delivery without using additional lipid-based transfection agent. We aimed to impart self-delivery property to a non-viral vector by using the cell and nucleus penetrating properties of YopM proteins from the three Yersinia spp. (Y. pestis, Y. enterocolotica and Y. pseudotuberculosis). Plasmid DNA (pDNA) encoding green fluorescent protein (GFP) was labeled with quantum dots (QDs) via peptide-nucleic acid (PNA) recognition site. Recombinant YopM protein was then attached to the conjugate via a second PNA recognition site. The YopM ̶ QDs ̶ pDNA conjugate was transfected into HeLa cells without using additional transfection reagent. All three conjugates produced GFP fluorescence, indicating that the plasmid was successfully delivered to the nucleus. As control, naked pDNA was transfected into the cells by using a commercial transfection reagent. The Y. pseudotuberculosis YopM-functionalized conjugate achieved the highest GFP expression, compared to other two YopM proteins and the transfection reagent. To the best of our knowledge, YopM protein was used for the first time in a non-viral gene delivery vector.

非病毒基因递送系统在安全有效的基因治疗方面具有巨大潜力，而低效率的细胞和核吸收仍然是主要障碍。需要新颖的方法来增强非病毒载体的转染效率。根据这一需要，本研究的目的是构建一种无需使用其他基于脂质的转染剂即可实现基因传递的非病毒载体。我们的目标是通过使用来自三个耶尔森氏菌属的YopM蛋白的细胞和细胞核穿透特性，将自身传递特性赋予非病毒载体。（鼠疫耶尔森氏菌，小肠结肠炎耶尔森氏菌和假结核耶尔森氏菌）。通过肽核酸（PNA）识别位点，用量子点（QD）标记编码绿色荧光蛋白（GFP）的质粒DNA（pDNA）。然后，通过第二个PNA识别位点将重组YopM蛋白连接到缀合物上。将YopM QDs pDNA偶联物转染到HeLa细胞中，无需使用其他转染试剂。所有三种结合物均产生GFP荧光，表明该质粒已成功递送至细胞核。作为对照，通过使用商业转染试剂将裸露的pDNA转染到细胞中。该假结核菌与其他两种YopM蛋白和转染试剂相比，YopM功能化的偶联物实现了最高的GFP表达。据我们所知，YopM蛋白首次用于非病毒基因递送载体。