한빛사논문
- 조회1,790
Abstract
Hyunseo Koo†, Ikbum Park‡, Yoonhee Lee†, Hyun Jin Kim§, Jung Hoon Jung§, Joo Han Lee§, Youngkyu Kim*†, Joung-Hun Kim*§, and Joon Won Park*†
†Department of Chemistry, ‡Division of Integrative Biosciences and Biotechnology, and §Department of Life Sciences, Pohang University of Science and Technology, 77 Cheongam-Ro, Nam-Gu, Pohang 37673, Korea
*Corresponding Authors
Author Contributions
H. Koo and I. Park contributed equally to this work.
Abstract
MicroRNAs (miRNAs) play critical roles in controlling various cellular processes, and the expression levels of individual miRNAs can be considerably altered in pathological conditions such as cancer. Accurate quantification of miRNA at the single-cell level will lead to a better understanding of miRNA function. Here, we present a direct and sensitive method for miRNA detection using atomic force microscopy (AFM). A hybrid binding domain (HBD)-tethered tip enabled mature miRNAs, but not premature miRNAs, to be located individually on an adhesion force map. By scanning several sections of a micrometer-sized DNA spot, we were able to quantify the copy number of miR-134 in a single neuron and demonstrate that the expression was increased upon cell activation. Moreover, we visualized individual miR-134s on fixed neurons after membrane removal and observed 2-4 miR-134s in the area of 1.0 × 1.0 μm2 of soma. The number increased to 8-4 in stimulated neurons, and this change matches the ensemble-averaged increase in copy number. These findings indicate that miRNAs can be reliably quantified at the single cell level with AFM and that their distribution can be mapped at nanometric lateral resolution without modification or amplification. Furthermore, the analysis of miRNAs, mRNAs, and proteins in the same sample or region by scanning sequentially with different AFM tips would let us accurately understand the post-transcriptional regulation of biological processes.
논문정보
- Research article
- 2016년 08월 (BRIC 등록일 2016-09-05)
- 국내 연구진
- 바이오·의료융합 > 바이오센싱 및 나노바이오물질
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