Generation of a Telomere-Based Episomal Vector

We have developed a telomere‐based episome by large‐scale amplification in Escherichia colicells. This episome consists of a PAC vector in which a 6 Kb sequence, containing an array of telomeric repeats spaced by a synthetic sequence, is tandemly repeated by large‐scale multimerization in E. coli. A...

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Bibliographic Details
Published in:Biotechnology progress 2003-11, Vol.19 (6), p.1775-1780
Main Authors: d'Aiuto, Leonardo, Heras, Jose I. de las, Ross, Andrew, Shen, Ming Hong, Cooke, Howard
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Cell Line, Tumor
Chromosomes, Artificial, P1 Bacteriophage - genetics
Chromosomes, Artificial, P1 Bacteriophage - ultrastructure
Cloning, Molecular - methods
episomes
Escherichia coli
Fibrosarcoma - genetics
Fibrosarcoma - pathology
Fundamental and applied biological sciences. Psychology
Gene Transfer Techniques
Genetic Vectors - genetics
Humans
Plasmids - genetics
Plasmids - ultrastructure
Telomere - genetics
Telomere - ultrastructure
Transfection - methods
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Summary:We have developed a telomere‐based episome by large‐scale amplification in Escherichia colicells. This episome consists of a PAC vector in which a 6 Kb sequence, containing an array of telomeric repeats spaced by a synthetic sequence, is tandemly repeated by large‐scale multimerization in E. coli. After transfection in human HT1080 cells, the construct, called clone 106, was able to persist in episomal form or integrated into some endogenous chromosomes. Integrations occurred exclusively at the telomeres. Episomes were still present in HT1080 cells after more than 100 days in the absence of selection. Integrations of clone 106 into the telomeric regions were retained only under selective conditions, and when the selection was removed the construct was progressively eliminated from the chromosome. The long‐term maintenance of clone 106 into human cells as an episome and its ability to integrate transiently into the telomeres of the host chromosomes suggest that this PAC‐based episome is potentially a good candidate vector for gene therapy applications.
ISSN:8756-7938
1520-6033
DOI:10.1021/bp0341500