title: Bioorthogonal tetrazine-mediated transfer reactions facilitate reaction turnover in nucleic acid-templated detection of microRNA.
abstract: Tetrazine ligations have proven to be a powerful bioorthogonal technique for the detection of many labeled biomolecules, but the ligating nature of these reactions can limit reaction turnover in templated chemistry. We have developed a transfer reaction between 7-azabenzonorbornadiene derivatives and fluorogenic tetrazines that facilitates turnover amplification of the fluorogenic response in nucleic acid-templated reactions. Fluorogenic tetrazine-mediated transfer (TMT) reaction probes can be used to detect DNA and microRNA (miRNA) templates to 0.5 and 5 pM concentrations, respectively. The endogenous oncogenic miRNA target mir-21 could be detected in crude cell lysates and detected by imaging in live cells. Remarkably, the technique is also able to differentiate between miRNA templates bearing a single mismatch with high signal to background. We imagine that TMT reactions could find wide application for amplified fluorescent detection of clinically relevant nucleic acid templates.
authors.affiliation: Department of Chemistry and Biochemistry, University of California, San Diego , La Jolla, California 92093, United States.
authors.forename: Haoxing
authors.initials: H
authors.lastname: Wu
authors.forename: Brandon T
authors.initials: BT
authors.lastname: Cisneros
authors.forename: Christian M
authors.initials: CM
authors.lastname: Cole
authors.forename: Neal K
authors.initials: NK
authors.lastname: Devaraj
mesh_terms: D001372:Aza Compounds / Q000737:chemistry; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006573:Heterocyclic Compounds, 1-Ring / Q000737:chemistry*; D006801:Humans; D061986:MCF-7 Cells; D035683:MicroRNAs / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D008958:Models, Molecular; D009690:Nucleic Acid Conformation
chemical_list: D001372:Aza Compounds; D005456:Fluorescent Dyes; D006573:Heterocyclic Compounds, 1-Ring; D035683:MicroRNAs