An in vivo reporter system for measuring increased inclusion of exon 7 in SMN2 mRNA

Potential therapy of SMA

M. L. Zhang, C. L. Lorson, Elliot Androphy, J. Zhou

Research output: Contribution to journalArticle

97 Citations (Scopus)

Abstract

Spinal muscular atrophy (SMA) is a degenerative motor neuron disorder resulting from homozygous loss of the SMN1 gene. SMN2, a nearly identical copy gene, is preserved in SMA patients. A single nucleotide difference between SMN1 and SMN2 causes exon 7 skipping in the majority of SMN2 mRNA. Gene therapy through modulation of SMN2 gene transcription in SMA patients may be possible. We constructed a series of SMN mini-genes comprised of SMN exon 6 to exon 8 sequences fused to green fluorescence protein (GFP) or luciferase reporters, to monitor SMN exon 7 splicing. These reporters recapitulated the splicing patterns of the endogenous SMN gene in stable cell lines. The SMN1-luciferase reporter was approximately 3.5-fold more active than SMN2-luciferase and SMN1-GFP intensities were visually distinguishable from SMN2-GFP. We have screened chemical inducers and inhibitors of kinase pathways using stable SMN-reporter lines and found that the phosphatase inhibitor sodium vanadate specifically stimulated exon 7 inclusion within SMN2 mRNAs. This is the first compound identified that can stimulate exon 7 inclusion into transcripts derived from the endogenous SMN2 gene. These results demonstrate that this system can be utilized to identify small molecules that regulate the splicing of SMN exon 7.

Original languageEnglish (US)
Pages (from-to)1532-1538
Number of pages7
JournalGene Therapy
Volume8
Issue number20
DOIs
StatePublished - 2001
Externally publishedYes

Fingerprint

Spinal Muscular Atrophy
Exons
Messenger RNA
Luciferases
Genes
Fluorescence
Therapeutics
Proteins
Vanadates
Motor Neurons
Phosphoric Monoester Hydrolases
Genetic Therapy
Phosphotransferases
Nucleotides
Cell Line

Keywords

  • Gene therapy
  • High throughput screening (HTS)
  • Small molecules
  • Spinal muscular atrophy (SMA)
  • Splicing
  • Survival motor neuron (SMN)

ASJC Scopus subject areas

  • Genetics

Cite this

An in vivo reporter system for measuring increased inclusion of exon 7 in SMN2 mRNA : Potential therapy of SMA. / Zhang, M. L.; Lorson, C. L.; Androphy, Elliot; Zhou, J.

In: Gene Therapy, Vol. 8, No. 20, 2001, p. 1532-1538.

Research output: Contribution to journalArticle

@article{29abe9eca0204b0e969b69acf2521752,
title = "An in vivo reporter system for measuring increased inclusion of exon 7 in SMN2 mRNA: Potential therapy of SMA",
abstract = "Spinal muscular atrophy (SMA) is a degenerative motor neuron disorder resulting from homozygous loss of the SMN1 gene. SMN2, a nearly identical copy gene, is preserved in SMA patients. A single nucleotide difference between SMN1 and SMN2 causes exon 7 skipping in the majority of SMN2 mRNA. Gene therapy through modulation of SMN2 gene transcription in SMA patients may be possible. We constructed a series of SMN mini-genes comprised of SMN exon 6 to exon 8 sequences fused to green fluorescence protein (GFP) or luciferase reporters, to monitor SMN exon 7 splicing. These reporters recapitulated the splicing patterns of the endogenous SMN gene in stable cell lines. The SMN1-luciferase reporter was approximately 3.5-fold more active than SMN2-luciferase and SMN1-GFP intensities were visually distinguishable from SMN2-GFP. We have screened chemical inducers and inhibitors of kinase pathways using stable SMN-reporter lines and found that the phosphatase inhibitor sodium vanadate specifically stimulated exon 7 inclusion within SMN2 mRNAs. This is the first compound identified that can stimulate exon 7 inclusion into transcripts derived from the endogenous SMN2 gene. These results demonstrate that this system can be utilized to identify small molecules that regulate the splicing of SMN exon 7.",
keywords = "Gene therapy, High throughput screening (HTS), Small molecules, Spinal muscular atrophy (SMA), Splicing, Survival motor neuron (SMN)",
author = "Zhang, {M. L.} and Lorson, {C. L.} and Elliot Androphy and J. Zhou",
year = "2001",
doi = "10.1038/sj.gt.3301550",
language = "English (US)",
volume = "8",
pages = "1532--1538",
journal = "Gene Therapy",
issn = "0969-7128",
publisher = "Nature Publishing Group",
number = "20",

}

TY - JOUR

T1 - An in vivo reporter system for measuring increased inclusion of exon 7 in SMN2 mRNA

T2 - Potential therapy of SMA

AU - Zhang, M. L.

AU - Lorson, C. L.

AU - Androphy, Elliot

AU - Zhou, J.

PY - 2001

Y1 - 2001

N2 - Spinal muscular atrophy (SMA) is a degenerative motor neuron disorder resulting from homozygous loss of the SMN1 gene. SMN2, a nearly identical copy gene, is preserved in SMA patients. A single nucleotide difference between SMN1 and SMN2 causes exon 7 skipping in the majority of SMN2 mRNA. Gene therapy through modulation of SMN2 gene transcription in SMA patients may be possible. We constructed a series of SMN mini-genes comprised of SMN exon 6 to exon 8 sequences fused to green fluorescence protein (GFP) or luciferase reporters, to monitor SMN exon 7 splicing. These reporters recapitulated the splicing patterns of the endogenous SMN gene in stable cell lines. The SMN1-luciferase reporter was approximately 3.5-fold more active than SMN2-luciferase and SMN1-GFP intensities were visually distinguishable from SMN2-GFP. We have screened chemical inducers and inhibitors of kinase pathways using stable SMN-reporter lines and found that the phosphatase inhibitor sodium vanadate specifically stimulated exon 7 inclusion within SMN2 mRNAs. This is the first compound identified that can stimulate exon 7 inclusion into transcripts derived from the endogenous SMN2 gene. These results demonstrate that this system can be utilized to identify small molecules that regulate the splicing of SMN exon 7.

AB - Spinal muscular atrophy (SMA) is a degenerative motor neuron disorder resulting from homozygous loss of the SMN1 gene. SMN2, a nearly identical copy gene, is preserved in SMA patients. A single nucleotide difference between SMN1 and SMN2 causes exon 7 skipping in the majority of SMN2 mRNA. Gene therapy through modulation of SMN2 gene transcription in SMA patients may be possible. We constructed a series of SMN mini-genes comprised of SMN exon 6 to exon 8 sequences fused to green fluorescence protein (GFP) or luciferase reporters, to monitor SMN exon 7 splicing. These reporters recapitulated the splicing patterns of the endogenous SMN gene in stable cell lines. The SMN1-luciferase reporter was approximately 3.5-fold more active than SMN2-luciferase and SMN1-GFP intensities were visually distinguishable from SMN2-GFP. We have screened chemical inducers and inhibitors of kinase pathways using stable SMN-reporter lines and found that the phosphatase inhibitor sodium vanadate specifically stimulated exon 7 inclusion within SMN2 mRNAs. This is the first compound identified that can stimulate exon 7 inclusion into transcripts derived from the endogenous SMN2 gene. These results demonstrate that this system can be utilized to identify small molecules that regulate the splicing of SMN exon 7.

KW - Gene therapy

KW - High throughput screening (HTS)

KW - Small molecules

KW - Spinal muscular atrophy (SMA)

KW - Splicing

KW - Survival motor neuron (SMN)

UR - http://www.scopus.com/inward/record.url?scp=0035158371&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035158371&partnerID=8YFLogxK

U2 - 10.1038/sj.gt.3301550

DO - 10.1038/sj.gt.3301550

M3 - Article

VL - 8

SP - 1532

EP - 1538

JO - Gene Therapy

JF - Gene Therapy

SN - 0969-7128

IS - 20

ER -