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The use of an adeno-associated viral vector for efficient bicistronic expression of two genes in the central nervous system

Research output: Chapter in Book/Report/Conference proceedingChapter

Standard

The use of an adeno-associated viral vector for efficient bicistronic expression of two genes in the central nervous system. / Hutson, Thomas Haynes; Kathe, Claudia; Menezes, Sean Christopher; Rooney, Marie-Claire; Bueler, Hansruedi; Moon, Lawrence David Falcon.

Axon Growth and Regeneration: Methods and Protocols. ed. / Andrew J. Murray. Vol. N/A N/A. ed. N/A : Springer New York, 2014. p. 189-207 (Methods in Molecular Biology; Vol. 1162).

Research output: Chapter in Book/Report/Conference proceedingChapter

Harvard

Hutson, TH, Kathe, C, Menezes, SC, Rooney, M-C, Bueler, H & Moon, LDF 2014, The use of an adeno-associated viral vector for efficient bicistronic expression of two genes in the central nervous system. in AJ Murray (ed.), Axon Growth and Regeneration: Methods and Protocols. N/A edn, vol. N/A, Methods in Molecular Biology, vol. 1162, Springer New York, N/A, pp. 189-207. https://doi.org/10.1007/978-1-4939-0777-9_16

APA

Hutson, T. H., Kathe, C., Menezes, S. C., Rooney, M-C., Bueler, H., & Moon, L. D. F. (2014). The use of an adeno-associated viral vector for efficient bicistronic expression of two genes in the central nervous system. In A. J. Murray (Ed.), Axon Growth and Regeneration: Methods and Protocols (N/A ed., Vol. N/A, pp. 189-207). (Methods in Molecular Biology; Vol. 1162). Springer New York. https://doi.org/10.1007/978-1-4939-0777-9_16

Vancouver

Hutson TH, Kathe C, Menezes SC, Rooney M-C, Bueler H, Moon LDF. The use of an adeno-associated viral vector for efficient bicistronic expression of two genes in the central nervous system. In Murray AJ, editor, Axon Growth and Regeneration: Methods and Protocols. N/A ed. Vol. N/A. N/A: Springer New York. 2014. p. 189-207. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-0777-9_16

Author

Hutson, Thomas Haynes ; Kathe, Claudia ; Menezes, Sean Christopher ; Rooney, Marie-Claire ; Bueler, Hansruedi ; Moon, Lawrence David Falcon. / The use of an adeno-associated viral vector for efficient bicistronic expression of two genes in the central nervous system. Axon Growth and Regeneration: Methods and Protocols. editor / Andrew J. Murray. Vol. N/A N/A. ed. N/A : Springer New York, 2014. pp. 189-207 (Methods in Molecular Biology).

Bibtex Download

@inbook{d863691c3f39417c8a85c592efda0b46,
title = "The use of an adeno-associated viral vector for efficient bicistronic expression of two genes in the central nervous system",
abstract = "Recombinant adeno-associated viral (AAV) vectors are one of the most promising therapeutic delivery systems for gene therapy to the central nervous system (CNS). Preclinical testing of novel gene therapies requires the careful design and production of AAV vectors and their successful application in a model of CNS injury. One major limitation of AAV vectors is their limited packaging capacity (<5 kb) making the co-expression of two genes (e.g., from two promoters) difficult. An internal ribosomal entry site has been used to express two genes: However, the second transgene is often expressed at lower levels than the first. In addition to this, achieving high levels of transduction in the CNS can be challenging. In this chapter we describe the cloning of a bicistronic AAV vector that uses the foot-and-mouth disease virus 2A sequence to efficiently express two genes from a single promoter. Bicistronic expression of a therapeutic gene and a reporter gene is desirable so that the axons from transduced neurons can be tracked and, after CNS injury, the amount of axonal sprouting or regeneration quantified. We go on to describe how to perform a pyramidotomy model of CNS injury and the injection of AAV vectors into the sensorimotor cortex to provide efficient transduction and bicistronic gene expression in cortical neurons such that transduced axons are detectable in the dorsal columns of the spinal cord.",
author = "Hutson, {Thomas Haynes} and Claudia Kathe and Menezes, {Sean Christopher} and Marie-Claire Rooney and Hansruedi Bueler and Moon, {Lawrence David Falcon}",
year = "2014",
doi = "10.1007/978-1-4939-0777-9_16",
language = "English",
isbn = "9781493907762",
volume = "N/A",
series = "Methods in Molecular Biology",
publisher = "Springer New York",
pages = "189--207",
editor = "Murray, {Andrew J.}",
booktitle = "Axon Growth and Regeneration",
edition = "N/A",

}

RIS (suitable for import to EndNote) Download

TY - CHAP

T1 - The use of an adeno-associated viral vector for efficient bicistronic expression of two genes in the central nervous system

AU - Hutson, Thomas Haynes

AU - Kathe, Claudia

AU - Menezes, Sean Christopher

AU - Rooney, Marie-Claire

AU - Bueler, Hansruedi

AU - Moon, Lawrence David Falcon

PY - 2014

Y1 - 2014

N2 - Recombinant adeno-associated viral (AAV) vectors are one of the most promising therapeutic delivery systems for gene therapy to the central nervous system (CNS). Preclinical testing of novel gene therapies requires the careful design and production of AAV vectors and their successful application in a model of CNS injury. One major limitation of AAV vectors is their limited packaging capacity (<5 kb) making the co-expression of two genes (e.g., from two promoters) difficult. An internal ribosomal entry site has been used to express two genes: However, the second transgene is often expressed at lower levels than the first. In addition to this, achieving high levels of transduction in the CNS can be challenging. In this chapter we describe the cloning of a bicistronic AAV vector that uses the foot-and-mouth disease virus 2A sequence to efficiently express two genes from a single promoter. Bicistronic expression of a therapeutic gene and a reporter gene is desirable so that the axons from transduced neurons can be tracked and, after CNS injury, the amount of axonal sprouting or regeneration quantified. We go on to describe how to perform a pyramidotomy model of CNS injury and the injection of AAV vectors into the sensorimotor cortex to provide efficient transduction and bicistronic gene expression in cortical neurons such that transduced axons are detectable in the dorsal columns of the spinal cord.

AB - Recombinant adeno-associated viral (AAV) vectors are one of the most promising therapeutic delivery systems for gene therapy to the central nervous system (CNS). Preclinical testing of novel gene therapies requires the careful design and production of AAV vectors and their successful application in a model of CNS injury. One major limitation of AAV vectors is their limited packaging capacity (<5 kb) making the co-expression of two genes (e.g., from two promoters) difficult. An internal ribosomal entry site has been used to express two genes: However, the second transgene is often expressed at lower levels than the first. In addition to this, achieving high levels of transduction in the CNS can be challenging. In this chapter we describe the cloning of a bicistronic AAV vector that uses the foot-and-mouth disease virus 2A sequence to efficiently express two genes from a single promoter. Bicistronic expression of a therapeutic gene and a reporter gene is desirable so that the axons from transduced neurons can be tracked and, after CNS injury, the amount of axonal sprouting or regeneration quantified. We go on to describe how to perform a pyramidotomy model of CNS injury and the injection of AAV vectors into the sensorimotor cortex to provide efficient transduction and bicistronic gene expression in cortical neurons such that transduced axons are detectable in the dorsal columns of the spinal cord.

U2 - 10.1007/978-1-4939-0777-9_16

DO - 10.1007/978-1-4939-0777-9_16

M3 - Chapter

C2 - 24838969

SN - 9781493907762

VL - N/A

T3 - Methods in Molecular Biology

SP - 189

EP - 207

BT - Axon Growth and Regeneration

A2 - Murray, Andrew J.

PB - Springer New York

CY - N/A

ER -

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