Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/84347
Title: GapmeR cellular internalization by macropinocytosis induces sequence-specific gene silencing in human primary T-cells
Authors: Fazil, Mobashar Hussain Urf Turabe
Ong, Seow Theng
Chalasani, Madhavi Latha Somaraju
Low, Jian Hui
Kizhakeyil, Atish
Mamidi, Akshay
Lim, Carey Fang Hui
Wright, Graham D.
Lakshminarayanan, Rajamani
Kelleher, Dermot
Verma, Navin Kumar
Keywords: T cells
Translational immunology
Issue Date: 2016
Source: Fazil, M. H. U. T., Ong, S. T., Chalasani, M. L. S., Low, J. H., Kizhakeyil, A., Mamidi, A., et al. (2016). GapmeR cellular internalization by macropinocytosis induces sequence-specific gene silencing in human primary T-cells. Scientific Reports, 6, 37721-.
Series/Report no.: Scientific Reports
Abstract: Post-transcriptional gene silencing holds great promise in discovery research for addressing intricate biological questions and as therapeutics. While various gene silencing approaches, such as siRNA and CRISPR-Cas9 techniques, are available, these cannot be effectively applied to “hard-to-transfect” primary T-lymphocytes. The locked nucleic acid-conjugated chimeric antisense oligonucleotide, called “GapmeR”, is an emerging new class of gene silencing molecule. Here, we show that GapmeR internalizes into human primary T-cells through macropinocytosis. Internalized GapmeR molecules can associate with SNX5-positive macropinosomes in T-cells, as detected by super-resolution microscopy. Utilizing the intrinsic self-internalizing capability of GapmeR, we demonstrate significant and specific depletion (>70%) of the expression of 5 different endogenous proteins with varying molecular weights (18 kDa Stathmin, 80 kDa PKCε, 180 kDa CD11a, 220 kDa Talin1 and 450 kDa CG-NAP/AKAP450) in human primary and cultured T-cells. Further functional analysis confirms CG-NAP and Stathmin as regulators of T-cell motility. Thus, in addition to screening, identifying or verifying critical roles of various proteins in T-cell functioning, this study provides novel opportunities to silence individual or multiple genes in a subset of purified human primary T-cells that would be exploited as future therapeutics.
URI: https://hdl.handle.net/10356/84347
http://hdl.handle.net/10220/41790
ISSN: 2045-2322
DOI: 10.1038/srep37721
Rights: © 2016 The Authors. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:LKCMedicine Journal Articles

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