Detecting and quantifying intracellular microRNAs(miRNAs)are a critical step in resolving a cancer diagnostic and resolving the ensemble of gene products that orchestrate the living state of cells.However,the nanoprob...Detecting and quantifying intracellular microRNAs(miRNAs)are a critical step in resolving a cancer diagnostic and resolving the ensemble of gene products that orchestrate the living state of cells.However,the nanoprobe for detecting low abundance miRNAs in cell cytosol is restricted by either the"one-to-one"signaltrigger model or di culty for cytosol delivery.To address these challenges,we designed a lightharvesting nanoantenna-based nanoprobe,which directs excitation energy to a single molecule to sensitively detect cytosolic miRNA.With light irradiation,the light-harvesting nanoantenna e ectively disrupted lysosomal structures by generationof reactive oxygen species,substantially achieved cytosol delivery.The nanoantenna containing>4000 donor dyes can e ciently transfer excitation energy to one or two acceptors with 99%e ciency,leading to unprecedented signal amplification and biosensing sensitivity.The designed nanoantenna can quantify cytosolic miR-210 at zeptomolar level.The fluorescence lifetime of the donor exhibited good relationship with miR-210 concentration in the range of 0.032 to 2.97 amol/ngRNA.The zeptomole sensitivity of nanoantenna provides accurate bioimaging of miR-210 both in multiple cell lines and in vivo assay,which creates a pathway for the creation of miRNA toolbox for quantitative epigenetics and personalized medicine.展开更多
基金supported by start-up fund of Washington State University。
文摘Detecting and quantifying intracellular microRNAs(miRNAs)are a critical step in resolving a cancer diagnostic and resolving the ensemble of gene products that orchestrate the living state of cells.However,the nanoprobe for detecting low abundance miRNAs in cell cytosol is restricted by either the"one-to-one"signaltrigger model or di culty for cytosol delivery.To address these challenges,we designed a lightharvesting nanoantenna-based nanoprobe,which directs excitation energy to a single molecule to sensitively detect cytosolic miRNA.With light irradiation,the light-harvesting nanoantenna e ectively disrupted lysosomal structures by generationof reactive oxygen species,substantially achieved cytosol delivery.The nanoantenna containing>4000 donor dyes can e ciently transfer excitation energy to one or two acceptors with 99%e ciency,leading to unprecedented signal amplification and biosensing sensitivity.The designed nanoantenna can quantify cytosolic miR-210 at zeptomolar level.The fluorescence lifetime of the donor exhibited good relationship with miR-210 concentration in the range of 0.032 to 2.97 amol/ngRNA.The zeptomole sensitivity of nanoantenna provides accurate bioimaging of miR-210 both in multiple cell lines and in vivo assay,which creates a pathway for the creation of miRNA toolbox for quantitative epigenetics and personalized medicine.
