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  • 02월 19일 23시 이후 : 초록수정 불가능, 일정확인 및 검색만 가능

제119회 대한화학회 학술발표회, 총회 및 기기전시회 Visualization and Quantification of miRNA in a Single cell Using Atomic Force Spectroscopy

2017년 3월 20일 17시 47분 33초
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목 11시 : 24분
한국다우케미칼 우수논문상 수상자 구두발표
저자 및
구현서, 박익범1, 이윤희, 김현진2, 정정훈2, 이주한2, 김영규*, 김정훈2,*, 박준원*
POSTECH 화학과, Korea
1POSTECH 융합생명공학부, Korea
2POSTECH 생명과학과, Korea
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 8–14 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.