|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 276, Issue 16, 13226-13230, April 20, 2001
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the Apolipoprotein (apo) E is a polymorphic
plasma protein, synthesized mainly by liver. Here, we evaluate whether
synthetic DNA-RNA oligonucleotides (chimeraplasts) can convert a
dysfunctional isoform, apoE2 (C
Gene Correction of the Apolipoprotein (Apo) E2 Phenotype to
Wild-type ApoE3 by in Situ Chimeraplasty*
§,,¶
Department of Medicine, Royal Free and
University College Medical School, London NW3 2PF and the
§ Department of Biochemistry, Royal Holloway
University of London, Surrey TW20 0EX, United Kingdom
T, R158C), which causes Type
III hyperlipidemia and premature atherosclerosis, into apoE3. First, we
treated recombinant Chinese hamster ovary cells stably secreting
apoE2 with a 68-mer apoE2 to apoE3 chimeraplast. About one-third of
apoE2 was converted to apoE3, and the repair was stable through 12 passages. Subcloning treated cells produced both apoE2 and apoE3
clones. Direct sequencing and reverse transcription polymerase chain
reaction confirmed the genotype, whereas phenotypic change was verified
by isoelectric focusing and immunoblotting of secreted proteins.
Second, we established that the APOE2 gene can be targeted
both in vivo, using transgenic mice overexpressing human
apoE2, and in chromosomal context, using cultured lymphocytes from a
patient homozygous for the 
2 allele. We conclude that
chimeraplasty has the potential to convert the apoE2 mutation in
patients with Type III hyperlipidemia to apoE3.
*
This work was supported in part by Project Grant PG/99032
and by Ph.D. Studentship FS/97054 (to A. D. T.) from the British Heart Foundation.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  V. T. Ciavatta, S. A. Padove, J. H. Boatright, and J. M. Nickerson Mouse Retina Has Oligonucleotide-Induced Gene Repair Activity Invest. Ophthalmol. Vis. Sci., July 1, 2005; 46(7): 2291 - 2299. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Bertoni, G. E. Morris, and T. A. Rando Strand bias in oligonucleotide-mediated dystrophin gene editing Hum. Mol. Genet., January 15, 2005; 14(2): 221 - 233. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Bertoni, C. Lau, and T. A. Rando Restoration of dystrophin expression in mdx muscle cells by chimeraplast-mediated exon skipping Hum. Mol. Genet., May 15, 2003; 12(10): 1087 - 1099. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Liu, S. Cheng, A. J. v. Brabant, and E. B. Kmiec Rad51p and Rad54p, but not Rad52p, elevate gene repair in Saccharomyces cerevisiae directed by modified single-stranded oligonucleotide vectors Nucleic Acids Res., July 1, 2002; 30(13): 2742 - 2750. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Liu, M. C. Rice, M. Drury, S. Cheng, H. Gamper, and E. B. Kmiec Strand Bias in Targeted Gene Repair Is Influenced by Transcriptional Activity Mol. Cell. Biol., June 1, 2002; 22(11): 3852 - 3863. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. D. Richardson, L. B. Augustin, B. T. Kren, and C. J. Steer Gene Repair and Transposon-Mediated Gene Therapy Stem Cells, March 1, 2002; 20(2): 105 - 118. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Metz, M. DiCola, T. Kurihara, A. Bailey, B. Frank, B. Roecklein, M. Blaese, and B. Dickey Mode of Action of RNA/DNA Oligonucleotides : Progress in the Development of Gene Repair as a Therapy for {alpha}1-Antitrypsin Deficiency Chest, March 1, 2002; 121(2007): 91S - 97S. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. D. Harris, I. R. Graham, S. Schepelmann, A. K. Stannard, M. L. Roberts, B. L. Hodges, V. Hill, A. Amalfitano, D. G. Hassall, J. S. Owen, et al. Acute regression of advanced and retardation of early aortic atheroma in immunocompetent apolipoprotein-E (apoE) deficient mice by administration of a second generation [E1-, E3-, polymerase-] adenovirus vector expressing human apoE Hum. Mol. Genet., January 1, 2002; 11(1): 43 - 58. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. K. Stannard, D. R. Riddell, S. M. Sacre, A. D. Tagalakis, C. Langer, A. von Eckardstein, P. Cullen, T. Athanasopoulos, G. Dickson, and J. S. Owen Cell-derived Apolipoprotein E (ApoE) Particles Inhibit Vascular Cell Adhesion Molecule-1 (VCAM-1) Expression in Human Endothelial Cells J. Biol. Chem., November 30, 2001; 276(49): 46011 - 46016. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Liu, M. C. Rice, and E. B. Kmiec In vivo gene repair of point and frameshift mutations directed by chimeric RNA/DNA oligonucleotides and modified single-stranded oligonucleotides Nucleic Acids Res., October 15, 2001; 29(20): 4238 - 4250. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |