A picornaviral 2A-like sequence-based tricistronic vector allowing for high-level therapeutic gene expression coupled to a dual-reporter system

Mark J. Osborn; Angela Panoskaltsis-Mortari; Ron T. McElmurry; Scott K. Bell; Dario A A Vignali; Martin D. Ryan; Andrew C. Wilber; R. Scott McIvor; Jakub Tolar; Bruce R. Blazar

doi:10.1016/j.ymthe.2005.04.013

A picornaviral 2A-like sequence-based tricistronic vector allowing for high-level therapeutic gene expression coupled to a dual-reporter system

Mark J. Osborn, Angela Panoskaltsis-Mortari, Ron T. McElmurry, Scott K. Bell, Dario A A Vignali, Martin D. Ryan, Andrew C. Wilber, R. Scott McIvor, Jakub Tolar, Bruce R. Blazar

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55 Scopus citations

Abstract

The 2A-like sequences from members of the picornavirus family were utilized to construct a tricistronic vector bearing the human iduronidase (IDUA) gene along with the firefly luciferase and DsRed2 reporter genes. The 2A-like sequences mediate a cotranslational cleavage event resulting in the release of each individual protein product. Efficient cleavage was observed and all three proteins were functional in vitro and in vivo, allowing for supratherapeutic IDUA enzyme levels and the coexpression of luciferase and DsRed2 expression, which enabled us to track gene expression.

Original language	English (US)
Pages (from-to)	569-574
Number of pages	6
Journal	Molecular Therapy
Volume	12
Issue number	3
DOIs	https://doi.org/10.1016/j.ymthe.2005.04.013
State	Published - Sep 2005

Bibliographical note

Funding Information:
The authors express their gratitude to Kevin Tram and Andrew Price for their technical assistance and advise. This work was supported in part by a University of Minnesota Children's Cancer Research Fund award (MJO) and NIH grants R01 HL49997 and HL52592 (BRB).

Keywords

Cis-acting hydrolase elements
Iduronidase
Internal ribosome entry site
Mucopolysaccharidosis type I
Multicistronic
Whole body imaging

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10.1016/j.ymthe.2005.04.013

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Osborn, M. J., Panoskaltsis-Mortari, A., McElmurry, R. T., Bell, S. K., Vignali, D. A. A., Ryan, M. D., Wilber, A. C., McIvor, R. S., Tolar, J., & Blazar, B. R. (2005). A picornaviral 2A-like sequence-based tricistronic vector allowing for high-level therapeutic gene expression coupled to a dual-reporter system. Molecular Therapy, 12(3), 569-574. https://doi.org/10.1016/j.ymthe.2005.04.013

Osborn, MJ , Panoskaltsis-Mortari, A, McElmurry, RT, Bell, SK, Vignali, DAA, Ryan, MD, Wilber, AC, McIvor, RS , Tolar, J & Blazar, BR 2005, 'A picornaviral 2A-like sequence-based tricistronic vector allowing for high-level therapeutic gene expression coupled to a dual-reporter system', Molecular Therapy, vol. 12, no. 3, pp. 569-574. https://doi.org/10.1016/j.ymthe.2005.04.013

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title = "A picornaviral 2A-like sequence-based tricistronic vector allowing for high-level therapeutic gene expression coupled to a dual-reporter system",

abstract = "The 2A-like sequences from members of the picornavirus family were utilized to construct a tricistronic vector bearing the human iduronidase (IDUA) gene along with the firefly luciferase and DsRed2 reporter genes. The 2A-like sequences mediate a cotranslational cleavage event resulting in the release of each individual protein product. Efficient cleavage was observed and all three proteins were functional in vitro and in vivo, allowing for supratherapeutic IDUA enzyme levels and the coexpression of luciferase and DsRed2 expression, which enabled us to track gene expression.",

keywords = "Cis-acting hydrolase elements, Iduronidase, Internal ribosome entry site, Mucopolysaccharidosis type I, Multicistronic, Whole body imaging",

author = "Osborn, {Mark J.} and Angela Panoskaltsis-Mortari and McElmurry, {Ron T.} and Bell, {Scott K.} and Vignali, {Dario A A} and Ryan, {Martin D.} and Wilber, {Andrew C.} and McIvor, {R. Scott} and Jakub Tolar and Blazar, {Bruce R.}",

note = "Funding Information: The authors express their gratitude to Kevin Tram and Andrew Price for their technical assistance and advise. This work was supported in part by a University of Minnesota Children's Cancer Research Fund award (MJO) and NIH grants R01 HL49997 and HL52592 (BRB).",

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doi = "10.1016/j.ymthe.2005.04.013",

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AU - Panoskaltsis-Mortari, Angela

AU - McElmurry, Ron T.

AU - Bell, Scott K.

AU - Vignali, Dario A A

AU - Ryan, Martin D.

AU - Wilber, Andrew C.

AU - McIvor, R. Scott

AU - Tolar, Jakub

AU - Blazar, Bruce R.

N1 - Funding Information: The authors express their gratitude to Kevin Tram and Andrew Price for their technical assistance and advise. This work was supported in part by a University of Minnesota Children's Cancer Research Fund award (MJO) and NIH grants R01 HL49997 and HL52592 (BRB).

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AB - The 2A-like sequences from members of the picornavirus family were utilized to construct a tricistronic vector bearing the human iduronidase (IDUA) gene along with the firefly luciferase and DsRed2 reporter genes. The 2A-like sequences mediate a cotranslational cleavage event resulting in the release of each individual protein product. Efficient cleavage was observed and all three proteins were functional in vitro and in vivo, allowing for supratherapeutic IDUA enzyme levels and the coexpression of luciferase and DsRed2 expression, which enabled us to track gene expression.

KW - Cis-acting hydrolase elements

KW - Iduronidase

KW - Internal ribosome entry site

KW - Mucopolysaccharidosis type I

KW - Multicistronic

KW - Whole body imaging

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A picornaviral 2A-like sequence-based tricistronic vector allowing for high-level therapeutic gene expression coupled to a dual-reporter system

Abstract

Bibliographical note

Keywords

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