title: Smart Magnetic Nanosensors Synthesized through Layer-by-Layer Deposition of Molecular Beacons for Noninvasive and Longitudinal Monitoring of Cellular mRNA.
abstract: Noninvasive and longitudinal monitoring of gene expression in living cells is essential for understanding and monitoring cellular activities. Herein, a smart magnetic nanosensor is constructed for the real-time, noninvasive, and longitudinal monitoring of cellular mRNA expression through the layer-by-layer deposition of molecular beacons (MBs) and polyethylenimine on the iron oxide nanoparticles. The loading of MBs, responsible for the signal intensity and the tracking time, was easily tuned with the number of layers incorporated. The idea was first demonstrated with the magnetic nanosensors for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA, which was efficiently internalized into the cells under the influence of magnetic field. This nanosensor allowed the continuous monitoring of the cellular GAPDH mRNA expression for 1 month. Then this platform was further utilized to incorporate two kinds of MBs for alkaline phosphatase (ALP) and GAPDH mRNAs, respectively. The multifunctional nanosensors permitted the simultaneous monitoring of the reference gene (GAPDH mRNA) and the early osteogenic differentiation marker (ALP mRNA) expression. When the fluorescence signal ratio between ALP mRNA MBs and GAPDH mRNA MBs was taken, the dynamic osteogenic differentiation process of MSCs was accurately monitored.
authors.affiliation: School of Chemical and Biomedical Engineering, Nanyang Technological University , 70 Nanyang Drive, Singapore 637457, Singapore.
authors.forename: Min
authors.initials: M
authors.lastname: Wang
authors.affiliation: School of Chemical and Biomedical Engineering, Nanyang Technological University , 70 Nanyang Drive, Singapore 637457, Singapore.|Key Laboratory for Organic Electronics and Information Displays, Nanjing University of Posts and Telecommunications , Nanjing 210046, China.
authors.forename: Xiaochun
authors.initials: X
authors.lastname: Hou
authors.affiliation: School of Chemical and Biomedical Engineering, Nanyang Technological University , 70 Nanyang Drive, Singapore 637457, Singapore.
authors.forename: Christian
authors.initials: C
authors.lastname: Wiraja
authors.forename: Libo
authors.initials: L
authors.lastname: Sun
authors.forename: Zhichuan J
authors.initials: ZJ
authors.lastname: Xu
authors.affiliation: School of Chemical and Biomedical Engineering, Nanyang Technological University , 70 Nanyang Drive, Singapore 637457, Singapore.
authors.forename: Chenjie
authors.initials: C
authors.lastname: Xu
mesh_terms: D000469:Alkaline Phosphatase / Q000235:genetics; D015374:Biosensing Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D000067493:Dynamic Light Scattering; D005290:Ferric Compounds / Q000737:chemistry; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006801:Humans; D058185:Magnetite Nanoparticles / Q000737:chemistry* / Q000633:toxicity / Q000648:ultrastructure; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D010012:Osteogenesis / Q000187:drug effects; D010316:Particle Size; D011094:Polyethyleneimine / Q000737:chemistry; D012333:RNA, Messenger / Q000032:analysis* / Q000378:metabolism
chemical_list: D005290:Ferric Compounds; D058185:Magnetite Nanoparticles; D012333:RNA, Messenger; C000499:ferric oxide; D011094:Polyethyleneimine; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D000469:Alkaline Phosphatase
keywords: layer-by-layer deposition; mRNA detection; magnetic nanoparticle; molecular beacon; nanosensor; osteogenic differentiation