title: Single-Stranded DNA Assisted Cell Penetrating Peptide-DNA Conjugation Strategy for Intracellular Imaging of Nucleases.
abstract: Cell penetrating peptides (CPPs) are very useful tools for delivery of DNA molecules into living cells without damaging the cell membranes. However, covalent conjugation of DNAs to CPPs is technically difficult, and the reactions between DNA and target nucleases are also liable to be affected by the cationic CPP molecules. In this work, we demonstrate that the electrostatic interactions between CPPs and single-stranded DNA (ssDNA) were stronger than those between CPP and double-stranded DNA (dsDNA). Taking advantage of this property, we developed an ssDNA protected CPP-DNA fluorescent probe which allowed for noninvasive and efficient cellular uptake and rapid imaging of target nucleases in living cells. The probe is highly sensitive and selective. This work represents the first example of using CPP-DNA conjugate to deliver DNA fluorescent probes for in situ imaging of nucleases within cells. The developed approach also holds great potential for the cellular delivery of other nucleic acid molecules for diagnosis or therapeutics purposes.
authors.affiliation: Beijing National Laboratory for Molecular Sciences, MOE Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University , Beijing 100871, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Chen
authors.affiliation: Beijing National Laboratory for Molecular Sciences, MOE Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University , Beijing 100871, China.
authors.forename: Simin
authors.initials: S
authors.lastname: Fang
authors.affiliation: Family Planning Research Institute/Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology , Wuhan, Hubei 430030, China.
authors.forename: Xianjin
authors.initials: X
authors.lastname: Xiao
authors.affiliation: Department of Chemistry, The Chinese University of Hong Kong , Shatin, New Territories, Hong Kong, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Zheng
authors.affiliation: Beijing National Laboratory for Molecular Sciences, MOE Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University , Beijing 100871, China.
authors.forename: Meiping
authors.initials: M
authors.lastname: Zhao
mesh_terms: D057846:Cell-Penetrating Peptides / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry*; D003851:Deoxyribonucleases / Q000032:analysis*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D061848:Optical Imaging; D055672:Static Electricity
chemical_list: D057846:Cell-Penetrating Peptides; D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D003851:Deoxyribonucleases