A gapmer aptamer nanobiosensor for real-time monitoring of transcription and translation in single cells

Shue Wang; Yuan Xiao; Donna D Zhang; Pak Kin Wong

doi:10.1016/j.biomaterials.2017.11.026

A gapmer aptamer nanobiosensor for real-time monitoring of transcription and translation in single cells

Biomaterials. 2018 Feb:156:56-64. doi: 10.1016/j.biomaterials.2017.11.026. Epub 2017 Nov 24.

Authors

Shue Wang¹, Yuan Xiao², Donna D Zhang³, Pak Kin Wong⁴

Affiliations

¹ Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI 48109, USA; Department of Aerospace and Mechanical Engineering, The University of Arizona, Tucson, AZ 85721, USA.
² Department of Aerospace and Mechanical Engineering, The University of Arizona, Tucson, AZ 85721, USA; Department of Biomedical Engineering, The Pennsylvania State University, University Park, PA 16802, USA.
³ Department of Pharmacology and Toxicology, The University of Arizona, Tucson, AZ 85721, USA.
⁴ Department of Aerospace and Mechanical Engineering, The University of Arizona, Tucson, AZ 85721, USA; Department of Biomedical Engineering, The Pennsylvania State University, University Park, PA 16802, USA; Department of Mechanical Engineering and Department of Surgery, The Pennsylvania State University, University Park, PA 16802, USA. Electronic address: pak@engr.psu.edu.

Abstract

Transcription and translation are under tight spatiotemporal regulation among cells to coordinate multicellular organization. Methods that allow massively parallel detection of gene expression dynamics at the single cell level are required for elucidating the complex regulatory mechanisms. Here we present a multiplex nanobiosensor for real-time monitoring of protein and mRNA expression dynamics in live cells based on gapmer aptamers and complementary locked nucleic acid probes. Using the multiplex nanobiosensor, we quantified spatiotemporal dynamics of vascular endothelial growth factor A mRNA and protein expressions in single human endothelial cells during microvascular self-organization. Our results revealed distinct gene regulatory processes in the heterogeneous cell subpopulations.

Keywords: Aptamer; Gapmer; Gene expression; Locked nucleic acid; Nanobiosensor.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Aptamers, Nucleotide / chemistry*
Biosensing Techniques / methods*
Computer Systems*
Gene Expression Profiling
Human Umbilical Vein Endothelial Cells / metabolism
Humans
Imaging, Three-Dimensional
Microvessels / metabolism
Nanoparticles / chemistry*
Phenotype
Protein Biosynthesis*
RNA, Messenger / genetics
RNA, Messenger / metabolism
Single-Cell Analysis*
Transcription, Genetic*
Vascular Endothelial Growth Factor A / genetics
Vascular Endothelial Growth Factor A / metabolism

Substances

Aptamers, Nucleotide
RNA, Messenger
Vascular Endothelial Growth Factor A

Abstract

Publication types

MeSH terms

Substances

Grants and funding