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Open AccessArticle

Internal Ribosome Entry Site Dramatically Reduces Transgene Expression in Hematopoietic Cells in a Position-Dependent Manner

by Qingyun Zheng 1,†, Xueyan Zhang 1,2,†, Hua Yang 2,3, Jinyan Xie 1, Yilin Xie 1, Jinzhong Chen 1,4, Chenghui Yu 1,2,4,* and Chen Zhong 1,*
1
State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200438, China
2
Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL 32610, USA
3
Department of Radiology, Central South University, Changsha, Hunan 410013, China
4
Yeda Research Institute of Gene and Cell Therapy, Taizhou, Zhejiang 318000, China
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Viruses 2019, 11(10), 920; https://doi.org/10.3390/v11100920
Received: 15 June 2019 / Revised: 21 September 2019 / Accepted: 6 October 2019 / Published: 8 October 2019
(This article belongs to the Special Issue Viruses and Cellular Metabolism)
Bicistronic transgene expression mediated by internal ribosome entry site (IRES) elements has been widely used. It co-expresses heterologous transgene products from a message RNA driven by a single promoter. Hematologic gene delivery is a promising treatment for both inherited and acquired diseases. A combined strategy was recently documented for potential genome editing in hematopoietic cells. A transduction efficiency exceeding ~90% can be achieved by capsid-optimized recombinant adeno-associated virus serotype 6 (rAAV6) vectors. In this study, to deliver an encephalomyocarditis virus (EMCV) IRES-containing rAAV6 genome into hematopoietic cells, we observed that EMCV IRES almost completely shut down the transgene expression during the process of mRNA–protein transition. In addition, position-dependent behavior was observed, in which only the EMCV IRES element located between a promoter and the transgenes had an inhibitory effect. Although further studies are warranted to evaluate the involvement of cellular translation machinery, our results propose the use of specific IRES elements or an alternative strategy, such as the 2A system, to achieve bicistronic transgene expression in hematopoietic cells. View Full-Text
Keywords: internal ribosome entry site (IRES); recombinant adeno-associated virus vector; hematopoietic cells; bicistronic transgene; gene therapy internal ribosome entry site (IRES); recombinant adeno-associated virus vector; hematopoietic cells; bicistronic transgene; gene therapy
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Zheng, Q.; Zhang, X.; Yang, H.; Xie, J.; Xie, Y.; Chen, J.; Yu, C.; Zhong, C. Internal Ribosome Entry Site Dramatically Reduces Transgene Expression in Hematopoietic Cells in a Position-Dependent Manner. Viruses 2019, 11, 920.

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