A Bimodal, Cationic, and Water-Soluble Calix[4]arene Conjugate: Design, Synthesis, Characterization, and Transfection of Red Fluorescent Protein Encoded Plasmid in Cancer Cells

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• 作者：Kushal Samanta ; Dnyanesh S. Ranade ; Aekta Upadhyay ; Prasad P. Kulkarni ; Chebrolu Pulla Rao
• 刊名：ACS Applied Materials & Interfaces
• 出版年：2017
• 出版时间：February 15, 2017
• 年：2017
• 卷：9
• 期：6
• 页码：5109-5117
• 全文大小：645K
• ISSN：1944-8252

文摘

A new bimodal fluorescent cationic calix[4]arene (L<sub>1sub>) conjugate has been synthesized in multiple steps and well characterized by NMR and electrospray ionization–mass spectrometry (ESI–MS) techniques. L<sub>1sub> has been investigated for its DNA binding ability by various spectroscopy techniques like absorption, fluorescence, and circular dichroism (CD). The formation of L<sub>1sub>–DNA complex has been confirmed by the gel electrophoresis in the presence of incremental concentration of L<sub>1sub>. To visualize the packing of the plasmid (pBR322), detailed tapping mode atomic force microscopy study has been performed, which revealed blob-like structure of plasmid upon addition of the incremental amount of L<sub>1sub>. Concentration dependent transfection ability of L<sub>1sub> has been established in MCF-7 cells by confocal microscopy by carrying the red fluorescent protein (RFP) encoded plasmid pCMV-tdTomato-N1 to emit both intrinsic fluorescence of L<sub>1sub> as well as that from RFP. All this has been possible in the absence of any adjuvant phospholipids (DOPE) that are commonly used as helper. Further transfection efficiency of L<sub>1sub> has been compared with the commercially available lipofectamine (LTX) in two cancer cell lines, MCF 7 and SH-SY5Y, and found that the L<sub>1sub> is as efficient as that of LTX. Hence, L<sub>1sub> is an efficient and effective cargo to transport genetic material into the cells.