Document Detail


Targeted beta-globin gene conversion in human hematopoietic CD34(+ )and Lin(-)CD38(-)cells.
MedLine Citation:
PMID:  11857070     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Chimeric oligonucleotides have been used successfully to correct point and frameshift mutations in several cell types, as well as in animal and plant models. However, their application to primitive human blood cells has been limited. In this investigation, chimeric oligonucleotides designed to direct a site-specific nucleotide exchange in the human beta-globin gene (an A to T substitution within codon 6) were introduced into normal human CD34(+) and Lin(-)CD38(-) cells via microinjection. This A to T nucleotide exchange introduces the single site mutation responsible for sickle cell anemia. In 23% of experimental samples, gene conversion was detected in the progeny of microinjected CD34(+) and Lin(-)CD38(-) cells that were cultured for at least 4 weeks. In addition, gene conversion was detected in the erythroid progeny of Lin(-)CD38(-) cells at the mRNA level. Conversion rates as high as 10-15% in 11% (five of 44) of experimental samples were confirmed by allele-specific PCR and sequence analysis of genomic DNA from the progeny of microinjected Lin(-)CD38(-) cells. Given that as few as 10% normal hematopoietic cells are sufficient to keep patients free of sickle cell disease, the level of conversion we have achieved in some samples may well be of therapeutic benefit in patients with sickle cell disease.
Authors:
H Liu; S Agarwal; E Kmiec; B R Davis
Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Gene therapy     Volume:  9     ISSN:  0969-7128     ISO Abbreviation:  Gene Ther.     Publication Date:  2002 Jan 
Date Detail:
Created Date:  2002-02-21     Completed Date:  2002-03-12     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  9421525     Medline TA:  Gene Ther     Country:  England    
Other Details:
Languages:  eng     Pagination:  118-26     Citation Subset:  IM    
Affiliation:
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA.
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MeSH Terms
Descriptor/Qualifier:
ADP-ribosyl Cyclase
Anemia, Sickle Cell / genetics*
Antigens, CD*
Antigens, CD34 / analysis
Antigens, CD38
Antigens, Differentiation / analysis
Cells, Cultured
Gene Conversion*
Gene Targeting / methods*
Gene Therapy / methods*
Globins / genetics*
Hematopoietic Stem Cells / pathology
Humans
Membrane Glycoproteins
Microinjections
NAD+ Nucleosidase / analysis
Point Mutation
Grant Support
ID/Acronym/Agency:
R01HL58583/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Antigens, CD; 0/Antigens, CD34; 0/Antigens, Differentiation; 0/Membrane Glycoproteins; 9004-22-2/Globins; EC 3.2.2.5/ADP-ribosyl Cyclase; EC 3.2.2.5/Antigens, CD38; EC 3.2.2.5/CD38 protein, human; EC 3.2.2.5/NAD+ Nucleosidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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