Real‐time imaging of dynamic cell reprogramming with nanosensors

Cellular reprogramming, the process by which somatic cells regain pluripotency, is relevant in many disease modeling, therapeutic, and drug discovery applications. Molecular evaluation of reprogramming (e.g., polymerase chain reaction, immunostaining) is typically disruptive, and only provides snaps...

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Main Authors: Wiraja, Christian, Yeo, David Chenloong, Tham, Khek-Chian, Chew, Sharon Wan Ting, Lim, Xinhong, Xu, Chenjie
Other Authors: School of Chemical and Biomedical Engineering
Format: Article
Language:English
Published: 2020
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Online Access:https://hdl.handle.net/10356/139022
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-1390222020-05-15T00:56:18Z Real‐time imaging of dynamic cell reprogramming with nanosensors Wiraja, Christian Yeo, David Chenloong Tham, Khek-Chian Chew, Sharon Wan Ting Lim, Xinhong Xu, Chenjie School of Chemical and Biomedical Engineering Interdisciplinary Graduate School (IGS) Lee Kong Chian School of Medicine (LKCMedicine) NTU Institute for Health Technologies NTU-Northwestern Institute for Nanomedicine Engineering::Chemical engineering Cell Reprogramming Molecular Beacons Cellular reprogramming, the process by which somatic cells regain pluripotency, is relevant in many disease modeling, therapeutic, and drug discovery applications. Molecular evaluation of reprogramming (e.g., polymerase chain reaction, immunostaining) is typically disruptive, and only provides snapshots of phenotypic traits. Gene reporter constructs facilitate live-cell evaluation but is labor intensive and may risk insertional mutagenesis during viral transfection. Herein, the utilization of a non-integrative nanosensor is demonstrated to visualize key reprogramming events in situ within live cells. Principally based on sustained intracellular release of encapsulated molecular probes, nanosensors successfully monitored mesenchymal-epithelial transition, pluripotency acquisition, and transdifferentiation events. Tracking the dynamic expression of four pivotal biomarkers (i.e., THY1, E-CADHERIN, OCT4, and GATA4 mRNA), nanosensor signal showed great agreement with polymerase chain reaction and gene reporter imaging (R2 > 0.9). Overall, such facile, versatile nanosensor enables real-time monitoring of low-frequency reprogramming events, thereby useful for high-throughput assessment, optimization, and biomarker-specific cell enrichment. 2020-05-15T00:56:18Z 2020-05-15T00:56:18Z 2018 Journal Article Wiraja, C., Yeo, D. C., Tham, K.-C., Chew, S. W. T., Lim, X., & Xu, C. (2018). Real‐time imaging of dynamic cell reprogramming with nanosensors. Small, 14(17), 1703440-. doi:10.1002/smll.201703440 1613-6810 https://hdl.handle.net/10356/139022 10.1002/smll.201703440 29611333 2-s2.0-85044713563 17 14 en Small © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. All rights reserved.
institution Nanyang Technological University
building NTU Library
country Singapore
collection DR-NTU
language English
topic Engineering::Chemical engineering
Cell Reprogramming
Molecular Beacons
spellingShingle Engineering::Chemical engineering
Cell Reprogramming
Molecular Beacons
Wiraja, Christian
Yeo, David Chenloong
Tham, Khek-Chian
Chew, Sharon Wan Ting
Lim, Xinhong
Xu, Chenjie
Real‐time imaging of dynamic cell reprogramming with nanosensors
description Cellular reprogramming, the process by which somatic cells regain pluripotency, is relevant in many disease modeling, therapeutic, and drug discovery applications. Molecular evaluation of reprogramming (e.g., polymerase chain reaction, immunostaining) is typically disruptive, and only provides snapshots of phenotypic traits. Gene reporter constructs facilitate live-cell evaluation but is labor intensive and may risk insertional mutagenesis during viral transfection. Herein, the utilization of a non-integrative nanosensor is demonstrated to visualize key reprogramming events in situ within live cells. Principally based on sustained intracellular release of encapsulated molecular probes, nanosensors successfully monitored mesenchymal-epithelial transition, pluripotency acquisition, and transdifferentiation events. Tracking the dynamic expression of four pivotal biomarkers (i.e., THY1, E-CADHERIN, OCT4, and GATA4 mRNA), nanosensor signal showed great agreement with polymerase chain reaction and gene reporter imaging (R2 > 0.9). Overall, such facile, versatile nanosensor enables real-time monitoring of low-frequency reprogramming events, thereby useful for high-throughput assessment, optimization, and biomarker-specific cell enrichment.
author2 School of Chemical and Biomedical Engineering
author_facet School of Chemical and Biomedical Engineering
Wiraja, Christian
Yeo, David Chenloong
Tham, Khek-Chian
Chew, Sharon Wan Ting
Lim, Xinhong
Xu, Chenjie
format Article
author Wiraja, Christian
Yeo, David Chenloong
Tham, Khek-Chian
Chew, Sharon Wan Ting
Lim, Xinhong
Xu, Chenjie
author_sort Wiraja, Christian
title Real‐time imaging of dynamic cell reprogramming with nanosensors
title_short Real‐time imaging of dynamic cell reprogramming with nanosensors
title_full Real‐time imaging of dynamic cell reprogramming with nanosensors
title_fullStr Real‐time imaging of dynamic cell reprogramming with nanosensors
title_full_unstemmed Real‐time imaging of dynamic cell reprogramming with nanosensors
title_sort real‐time imaging of dynamic cell reprogramming with nanosensors
publishDate 2020
url https://hdl.handle.net/10356/139022
_version_ 1681059516958900224