title: A Bimodal, Cationic, and Water-Soluble Calix[4]arene Conjugate: Design, Synthesis, Characterization, and Transfection of Red Fluorescent Protein Encoded Plasmid in Cancer Cells.
abstract: A new bimodal fluorescent cationic calix[4]arene (L1) conjugate has been synthesized in multiple steps and well characterized by NMR and electrospray ionization-mass spectrometry (ESI-MS) techniques. L1has been investigated for its DNA binding ability by various spectroscopy techniques like absorption, fluorescence, and circular dichroism (CD). The formation of L1-DNA complex has been confirmed by the gel electrophoresis in the presence of incremental concentration of L1. 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 L1. Concentration dependent transfection ability of L1has 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 L1as 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 L1has been compared with the commercially available lipofectamine (LTX) in two cancer cell lines, MCF 7 and SH-SY5Y, and found that the L1is as efficient as that of LTX. Hence, L1is an efficient and effective cargo to transport genetic material into the cells.
authors.affiliation: Bioinorganic Laboratory, Department of Chemistry, Indian Institute of Technology Bombay , Powai, Mumbai 400076, India.
authors.forename: Kushal
authors.initials: K
authors.lastname: Samanta
authors.affiliation: Bioprospecting Group, Agharkar Research Institute , G. G. Agarkar Road, Pune 411004, India.
authors.forename: Dnyanesh S
authors.initials: DS
authors.lastname: Ranade
authors.affiliation: Bioinorganic Laboratory, Department of Chemistry, Indian Institute of Technology Bombay , Powai, Mumbai 400076, India.
authors.forename: Aekta
authors.initials: A
authors.lastname: Upadhyay
authors.affiliation: Bioprospecting Group, Agharkar Research Institute , G. G. Agarkar Road, Pune 411004, India.
authors.forename: Prasad P
authors.initials: PP
authors.lastname: Kulkarni
authors.affiliation: Bioinorganic Laboratory, Department of Chemistry, Indian Institute of Technology Bombay , Powai, Mumbai 400076, India.
authors.forename: Chebrolu Pulla
authors.identifier: http://orcid.org/0000-0002-1004-0028
authors.initials: CP
authors.lastname: Rao
mesh_terms: D047250:Calixarenes; D006801:Humans; D008164:Luminescent Proteins; D009369:Neoplasms; D010636:Phenols; D010957:Plasmids; D014162:Transfection; D014867:Water; D000097573:Red Fluorescent Protein
chemical_list: D047250:Calixarenes; D008164:Luminescent Proteins; D010636:Phenols; D014867:Water; D000097573:Red Fluorescent Protein; C121325:calix(4)arene
keywords: Bimodal calix[4]arene conjugate; DNA binding; expression of RFP; plasmid condensation; transfection without DOPE