Imperial College London
Alternate

DrIsabelleSalles

Faculty of Medicine, Department of Immunology and Inflammation

Honorary Lecturer
 
 
 
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Contact

 

+44 (0)20 3313 2298i.salles

 
 
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Location

 

5S5Commonwealth BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Thijs:2015:10.1371/journal.pone.0132899,
author = {Thijs, T and Broos, K and Soenen, SJ and Vandenbulcke, A and Vanhoorelbeke, K and Deckmyn, H and Salles, I},
doi = {10.1371/journal.pone.0132899},
journal = {PLOS One},
title = {Artificial MiRNA Knockdown of Platelet Glycoprotein Ib alpha: A Tool for Platelet Gene Silencing},
url = {http://dx.doi.org/10.1371/journal.pone.0132899},
volume = {10},
year = {2015}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - In recent years, candidate genes and proteins implicated in platelet function have beenidentified by various genomic approaches. To elucidate their exact role, we aimed todevelop a method to apply miRNA interference in platelet progenitor cells by using GPIbαas a proof-of-concept target protein. After in silico and in vitro screening of siRNAs targetingGPIbα (siGPIBAs), we developed artificial miRNAs (miGPIBAs), which were tested inCHO cells stably expressing GPIb-IX complex and megakaryoblastic DAMI cells. Introductionof siGPIBAs in CHO GPIb-IX cells resulted in 44 to 75% and up to 80% knockdown ofGPIbα expression using single or combined siRNAs, respectively. Conversion of siGPIBAsto miGPIBAs resulted in reduced silencing efficiency, which could however be circumventedby tandem integration of two hairpins targeting different regions of GPIBA mRNAwhere 72% GPIbα knockdown was achieved. CHO GPIb-IX cells transfected with the miGPIBAconstruct displayed a significant decrease in their ability to aggregate characterizedby lower aggregate numbers and size compared to control CHO GPIb-IX cells. Moreimportantly, we successfully silenced GPIbα in differentiating megakaryoblastic DAMIcells that exhibited morphological changes associated with actin organization. In conclusion,we here report the successful use of miRNA technology to silence a platelet proteinin megakaryoblastic cells and demonstrate its usefulness in functional assays. Hence, webelieve that artificial miRNAs are suitable tools to unravel the role of a protein of interest instem cells, megakaryocytes and platelets, thereby expanding their application to novelfields of basic and translational research.
AU  - Thijs,T
AU  - Broos,K
AU  - Soenen,SJ
AU  - Vandenbulcke,A
AU  - Vanhoorelbeke,K
AU  - Deckmyn,H
AU  - Salles,I
DO  - 10.1371/journal.pone.0132899
PY  - 2015///
SN  - 1932-6203
TI  - Artificial MiRNA Knockdown of Platelet Glycoprotein Ib alpha: A Tool for Platelet Gene Silencing
T2  - PLOS One
UR  - http://dx.doi.org/10.1371/journal.pone.0132899
UR  - http://hdl.handle.net/10044/1/25984
VL  - 10
ER  -
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