title: Accurate quantification of microRNA via single strand displacement reaction on DNA origami motif.
abstract: DNA origami is an emerging technology that assembles hundreds of staple strands and one single-strand DNA into certain nanopattern. It has been widely used in various fields including detection of biological molecules such as DNA, RNA and proteins. MicroRNAs (miRNAs) play important roles in post-transcriptional gene repression as well as many other biological processes such as cell growth and differentiation. Alterations of miRNAs' expression contribute to many human diseases. However, it is still a challenge to quantitatively detect miRNAs by origami technology. In this study, we developed a novel approach based on streptavidin and quantum dots binding complex (STV-QDs) labeled single strand displacement reaction on DNA origami to quantitatively detect the concentration of miRNAs. We illustrated a linear relationship between the concentration of an exemplary miRNA as miRNA-133 and the STV-QDs hybridization efficiency; the results demonstrated that it is an accurate nano-scale miRNA quantifier motif. In addition, both symmetrical rectangular motif and asymmetrical China-map motif were tested. With significant linearity in both motifs, our experiments suggested that DNA Origami motif with arbitrary shape can be utilized in this method. Since this DNA origami-based method we developed owns the unique advantages of simple, time-and-material-saving, potentially multi-targets testing in one motif and relatively accurate for certain impurity samples as counted directly by atomic force microscopy rather than fluorescence signal detection, it may be widely used in quantification of miRNAs.
authors.affiliation: Bio-X Institutes, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education), Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhu
authors.forename: Xiaolu
authors.initials: X
authors.lastname: Feng
authors.forename: Jingyu
authors.initials: J
authors.lastname: Lou
authors.forename: Weidong
authors.initials: W
authors.lastname: Li
authors.forename: Sheng
authors.initials: S
authors.lastname: Li
authors.forename: Hongxin
authors.initials: H
authors.lastname: Zhu
authors.forename: Lun
authors.initials: L
authors.lastname: Yang
authors.forename: Aiping
authors.initials: A
authors.lastname: Zhang
authors.forename: Lin
authors.initials: L
authors.lastname: He
authors.forename: Can
authors.initials: C
authors.lastname: Li
mesh_terms: D020816:Amino Acid Motifs; D019921:Biotinylation; D002454:Cell Differentiation; D049109:Cell Proliferation; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D004277:DNA, Single-Stranded / Q000737:chemistry; D006801:Humans; D035683:MicroRNAs / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D036103:Nanotechnology; D009693:Nucleic Acid Hybridization; D045663:Quantum Dots; D012323:RNA Processing, Post-Transcriptional; D019809:Streptavidin / Q000737:chemistry
chemical_list: D004277:DNA, Single-Stranded; C548234:MIRN133 microRNA, human; D035683:MicroRNAs; D004247:DNA; D019809:Streptavidin