title: DNase-activatable fluorescence probes visualizing the degradation of exogenous DNA in living cells.
abstract: This work presents a method to visualize the degradation of exogenous DNA in living cells using a novel type of activatable fluorescence imaging probe. Deoxyribonuclease (DNase)-activatable fluorescence probes (DFProbes) are composed of double strands deoxyribonucleic acid (dsDNA) which is labeled with fluorophore (ROX or Cy3) and quencher on the end of one of its strands, and stained with SYBR Green I. In the absence of DNase, DFProbes produce the green fluorescence signal of SYBR Green I. In the presence of DNase, SYBR Green I is removed from the DFProbes and the labeled fluorophore is separated from the quencher owing to the degradation of DFProbes by DNase, resulting in the decrease of the green fluorescence signal and the occurrence of a red fluorescence signal due to fluorescence resonance energy transfer (FRET). DNase in biological samples was detected using DFProbes and the fluorescence imaging in living cells was performed using DFprobe-modified Au nanoparticles. The results show that DFProbes have good responses to DNase, and can clearly visualize the degradation of exogenous DNA in cells in real time. The well-designed probes might be useful in tracing the dynamic changes of exogenous DNA and nanocarriers in vitro and in vivo.
authors.affiliation: CAS Key Lab of Health Informatics, Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, Guangdong, PR China.
authors.forename: Ping
authors.initials: P
authors.lastname: Gong
authors.forename: Bihua
authors.initials: B
authors.lastname: Shi
authors.forename: Pengfei
authors.initials: P
authors.lastname: Zhang
authors.forename: Dehong
authors.initials: D
authors.lastname: Hu
authors.forename: Mingbin
authors.initials: M
authors.lastname: Zheng
authors.forename: Cuifang
authors.initials: C
authors.lastname: Zheng
authors.forename: Duyang
authors.initials: D
authors.lastname: Gao
authors.forename: Lintao
authors.initials: L
authors.lastname: Cai
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D019556:COS Cells; D058948:Cell Tracking / Q000295:instrumentation / Q000379:methods*; D002522:Chlorocebus aethiops; D003199:Computer Systems; D004247:DNA / Q000378:metabolism*; D053938:DNA Fragmentation; D003851:Deoxyribonucleases / Q000378:metabolism* / Q000502:physiology; D003959:Diamines; D005456:Fluorescent Dyes / Q000032:analysis / Q000378:metabolism* / Q000493:pharmacokinetics; D006367:HeLa Cells; D006801:Humans; D008954:Models, Biological; D009930:Organic Chemicals / Q000032:analysis / Q000378:metabolism / Q000493:pharmacokinetics; D011804:Quinolines; D014162:Transfection / Q000295:instrumentation / Q000379:methods; D019076:Transgenes / Q000235:genetics
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D004247:DNA; D003851:Deoxyribonucleases