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Title: RNA Structure Design Improves Activity and Specificity of trans-Splicing-Triggered Cell Death in a Suicide Gene Therapy Approach
Authors: Poddar, S. 
Loh, P.S. 
Ooi, Z.H.
Osman, F.
Eul, J.
Patzel, V. 
Keywords: hepatocellular carcinoma
rational RNA design
RNA trans-splicing
suicide gene therapy
Issue Date: 2018
Publisher: Cell Press
Citation: Poddar, S., Loh, P.S., Ooi, Z.H., Osman, F., Eul, J., Patzel, V. (2018). RNA Structure Design Improves Activity and Specificity of trans-Splicing-Triggered Cell Death in a Suicide Gene Therapy Approach. Molecular Therapy - Nucleic Acids 11 : 41-56. ScholarBank@NUS Repository.
Rights: Attribution-NonCommercial-NoDerivatives 4.0 International
Abstract: Spliceosome-mediated RNA trans-splicing enables correction or labeling of pre-mRNA, but therapeutic applications are hampered by issues related to the activity and target specificity of trans-splicing RNA (tsRNA). We employed computational RNA structure design to improve both on-target activity and specificity of tsRNA in a herpes simplex virus thymidine kinase/ganciclovir suicide gene therapy approach targeting alpha fetoprotein (AFP), a marker of hepatocellular carcinoma (HCC) or human papillomavirus type 16 (HPV-16) pre-mRNA. While unstructured, mismatched target binding domains significantly improved 3? exon replacement (3'ER), 5? exon replacement (5'ER) correlated with the thermodynamic stability of the tsRNA 3? end. Alternative on-target trans-splicing was found to be a prevalent event. The specificity of trans-splicing with the intended target splice site was improved 10-fold by designing tsRNA that harbors secondary target binding domains shielding alternative on-target and blinding off-target splicing events. Such rationally designed suicide RNAs efficiently triggered death of HPV-16-transduced or hepatoblastoma-derived human tissue culture cells without evidence for off-target cell killing. Highest cell death activities were observed with novel dual-targeting tsRNAs programmed for trans-splicing toward AFP and a second HCC pre-mRNA biomarker. Our observations suggest trans-splicing represents a promising approach to suicide gene therapy. © 2018 The Author(s)
Source Title: Molecular Therapy - Nucleic Acids
ISSN: 2162-2531
DOI: 10.1016/j.omtn.2018.01.006
Rights: Attribution-NonCommercial-NoDerivatives 4.0 International
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