Document Detail


Input DNA ratio determines copy number of the 33 kb Factor IX gene on de novo human artificial chromosomes.
MedLine Citation:
PMID:  18059371     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Human artificial chromosomes (ACs) are non-integrating vectors that may be useful for gene therapy. They assemble in cultured cells following transfection of human centromeric alpha -satellite DNA and segregate efficiently alongside the host genome. In the present study, a 33 kilobase (kb) Factor IX (FIX) gene was incorporated into mitotically stable ACs in human HT1080 lung derived cells using co-transfection of a bacterial artificial chromosome (BAC) harboring synthetic alpha -satellite DNA and a P1 artificial chromosome(PAC) that spans the FIX locus. ACs were detected in >or=90% of chromosome spreads in 8 of 19 lines expanded from drug resistant colonies. FIX transgene copy number on ACs was determined by input DNA transfection ratios. Furthermore, a low level of FIX transcription was detected from ACs with multiple transgenes but not from those incorporating a single transgene, suggesting that reducing transgene number may limit misexpression. Their potential to segregate cross species was measured by transferring ACs into mouse and hamster cell lines using microcell-mediated chromosome transfer. Lines were obtained where ACs segregated efficiently. The stable segregation of ACs in rodent cells suggests that it should be possible to develop animal models to test the capacity of ACs to rescue FIX deficiency.
Authors:
Amy M Breman; Camie M Steiner; Roger B Slee; Brenda R Grimes
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2007-12-04
Journal Detail:
Title:  Molecular therapy : the journal of the American Society of Gene Therapy     Volume:  16     ISSN:  1525-0024     ISO Abbreviation:  Mol. Ther.     Publication Date:  2008 Feb 
Date Detail:
Created Date:  2008-01-28     Completed Date:  2008-07-03     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100890581     Medline TA:  Mol Ther     Country:  United States    
Other Details:
Languages:  eng     Pagination:  315-23     Citation Subset:  IM    
Affiliation:
Department of Medical and Molecular Genetics, Indiana University School of Medicine, 975 W. Walnut Street, IB130, Indianapolis, Indiana, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Northern
Blotting, Southern
Cells, Cultured
Chromosomes, Artificial, Bacterial / genetics
Chromosomes, Artificial, Human / genetics*
Cricetinae
Cricetulus
Factor IX / genetics*
Fluorescent Antibody Technique
Humans
In Situ Hybridization, Fluorescence
Mice
Models, Genetic
Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
Transcription, Genetic / genetics
Transfection / methods
Transgenes / genetics
Chemical
Reg. No./Substance:
9001-28-9/Factor IX

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


Previous Document:  Induction of natural killer cell-dependent antitumor immunity by the Autographa californica multiple...
Next Document:  Downregulation of monocyte chemoattractant protein-1 involving short interfering RNA attenuates hapt...