Beta globin gene inhibition by antisense RNA transcripts

L. Xu, A. E. Ferry, C. Monteiro, B. S. Pace

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Sickle cell disease is caused by a mutation in the β globin gene leading to hemoglobin S (Hb S) production. Several approaches have been explored to prevent Hb S polymerization in red blood cells and the symptoms associated with this disorder. To this end we tested a mammalian expression vector carrying a human β globin antisense cDNA (pZeoβAS) fragment in a mouse erythroleukemia cell line expressing the human γ and β globin genes. We observed a relative reduction in β globin mRNA levels compared with γ mRNA levels in the presence of pZeoβAS. Moreover, analysis at the protein level showed an average 76% decrease in β chains and a 517% increase in γ chain biosynthesis. The inhibitory effect of the antisense vector on globin expression was maintained long term in culture. The expression vector pZeoβAS was also transfected into primary erythroid progenitors to test its effects on globin genes undergoing normal developmental switching during differentiation. We observed a relative reduction of β globin mRNA levels compared with γ mRNA levels. These results support a novel role for antisense cDNA expression vectors as an alternative gene therapy strategy to inhibit β(s) gene expression in sickle cell disease.

Original languageEnglish (US)
Pages (from-to)438-444
Number of pages7
JournalGene Therapy
Volume7
Issue number5
DOIs
StatePublished - Mar 2000

Fingerprint

Antisense RNA
beta-Globins
Globins
Genes
Sickle Hemoglobin
Messenger RNA
Sickle Cell Anemia
Complementary DNA
Leukemia, Erythroblastic, Acute
Complementary Therapies
Polymerization
Genetic Therapy
Erythrocytes
Gene Expression
Cell Line
Mutation

Keywords

  • Antisense cDNA
  • Expression vector
  • Oligodeoxynucleotide
  • β globin
  • γ globin

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Beta globin gene inhibition by antisense RNA transcripts. / Xu, L.; Ferry, A. E.; Monteiro, C.; Pace, B. S.

In: Gene Therapy, Vol. 7, No. 5, 03.2000, p. 438-444.

Research output: Contribution to journalArticle

Xu, L. ; Ferry, A. E. ; Monteiro, C. ; Pace, B. S. / Beta globin gene inhibition by antisense RNA transcripts. In: Gene Therapy. 2000 ; Vol. 7, No. 5. pp. 438-444.
@article{5c2cb976604f4f438e7e882d214dd6f2,
title = "Beta globin gene inhibition by antisense RNA transcripts",
abstract = "Sickle cell disease is caused by a mutation in the β globin gene leading to hemoglobin S (Hb S) production. Several approaches have been explored to prevent Hb S polymerization in red blood cells and the symptoms associated with this disorder. To this end we tested a mammalian expression vector carrying a human β globin antisense cDNA (pZeoβAS) fragment in a mouse erythroleukemia cell line expressing the human γ and β globin genes. We observed a relative reduction in β globin mRNA levels compared with γ mRNA levels in the presence of pZeoβAS. Moreover, analysis at the protein level showed an average 76{\%} decrease in β chains and a 517{\%} increase in γ chain biosynthesis. The inhibitory effect of the antisense vector on globin expression was maintained long term in culture. The expression vector pZeoβAS was also transfected into primary erythroid progenitors to test its effects on globin genes undergoing normal developmental switching during differentiation. We observed a relative reduction of β globin mRNA levels compared with γ mRNA levels. These results support a novel role for antisense cDNA expression vectors as an alternative gene therapy strategy to inhibit β(s) gene expression in sickle cell disease.",
keywords = "Antisense cDNA, Expression vector, Oligodeoxynucleotide, β globin, γ globin",
author = "L. Xu and Ferry, {A. E.} and C. Monteiro and Pace, {B. S.}",
year = "2000",
month = "3",
doi = "10.1038/sj.gt.3301106",
language = "English (US)",
volume = "7",
pages = "438--444",
journal = "Gene Therapy",
issn = "0969-7128",
publisher = "Nature Publishing Group",
number = "5",

}

TY - JOUR

T1 - Beta globin gene inhibition by antisense RNA transcripts

AU - Xu, L.

AU - Ferry, A. E.

AU - Monteiro, C.

AU - Pace, B. S.

PY - 2000/3

Y1 - 2000/3

N2 - Sickle cell disease is caused by a mutation in the β globin gene leading to hemoglobin S (Hb S) production. Several approaches have been explored to prevent Hb S polymerization in red blood cells and the symptoms associated with this disorder. To this end we tested a mammalian expression vector carrying a human β globin antisense cDNA (pZeoβAS) fragment in a mouse erythroleukemia cell line expressing the human γ and β globin genes. We observed a relative reduction in β globin mRNA levels compared with γ mRNA levels in the presence of pZeoβAS. Moreover, analysis at the protein level showed an average 76% decrease in β chains and a 517% increase in γ chain biosynthesis. The inhibitory effect of the antisense vector on globin expression was maintained long term in culture. The expression vector pZeoβAS was also transfected into primary erythroid progenitors to test its effects on globin genes undergoing normal developmental switching during differentiation. We observed a relative reduction of β globin mRNA levels compared with γ mRNA levels. These results support a novel role for antisense cDNA expression vectors as an alternative gene therapy strategy to inhibit β(s) gene expression in sickle cell disease.

AB - Sickle cell disease is caused by a mutation in the β globin gene leading to hemoglobin S (Hb S) production. Several approaches have been explored to prevent Hb S polymerization in red blood cells and the symptoms associated with this disorder. To this end we tested a mammalian expression vector carrying a human β globin antisense cDNA (pZeoβAS) fragment in a mouse erythroleukemia cell line expressing the human γ and β globin genes. We observed a relative reduction in β globin mRNA levels compared with γ mRNA levels in the presence of pZeoβAS. Moreover, analysis at the protein level showed an average 76% decrease in β chains and a 517% increase in γ chain biosynthesis. The inhibitory effect of the antisense vector on globin expression was maintained long term in culture. The expression vector pZeoβAS was also transfected into primary erythroid progenitors to test its effects on globin genes undergoing normal developmental switching during differentiation. We observed a relative reduction of β globin mRNA levels compared with γ mRNA levels. These results support a novel role for antisense cDNA expression vectors as an alternative gene therapy strategy to inhibit β(s) gene expression in sickle cell disease.

KW - Antisense cDNA

KW - Expression vector

KW - Oligodeoxynucleotide

KW - β globin

KW - γ globin

UR - http://www.scopus.com/inward/record.url?scp=0034114811&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034114811&partnerID=8YFLogxK

U2 - 10.1038/sj.gt.3301106

DO - 10.1038/sj.gt.3301106

M3 - Article

C2 - 10694826

AN - SCOPUS:0034114811

VL - 7

SP - 438

EP - 444

JO - Gene Therapy

JF - Gene Therapy

SN - 0969-7128

IS - 5

ER -