Mammalian Artificial Chromosomes: Prospects for Gene Therapy

2003 ◽  
pp. 165-178
Author(s):  
Tom A. Ebersole ◽  
Christine J. Farr
Keyword(s):  
Gene Therapy ◽  
Artificial Chromosomes

Implementing the human artificial chromosome gene therapy platform remains challenging, but continuous animal model research will advance the platform closer to clinical trials

2021 ◽  
Author(s):  
Moataz Dowaidar
Keyword(s):  
Gene Therapy ◽  
Clinical Trials ◽  
Animal Model ◽  
Human Artificial Chromosome ◽  
Target Cells ◽  
Minimal Risk ◽  
Artificial Chromosomes ◽  
Model Research ◽  
Alphoid Dna

A normal degree of ectopic gene expression, infinite retention in target cells without chromosomal integration, minimal risk of cell or neoplastic transformation, and minimal or no immunogenicity are all critical characteristics for vectors employed in gene therapy. HACs were produced and used as autonomous vectors to compensate for genetic defects in mouse and human cell cultures. Bottom-up human artificial chromosomes (HACs) were studied for functional transgene expression in vitro and in vivo mice models. The primary advantages of synthesized alphoid-HACs over top-down HACs are their defined and documented structure, as well as their relative simplicity of modification in adding numerous Cre-lox-type transgen loading sites. The HAC transfer method's efficacy has greatly increased in recent years. Despite significant progress in developing alphoid-HAC-based gene therapy models, the technology still has a number of drawbacks, including low HAC efficiency, complex repeated HAC alphoid-DNA structure, large DNA fragmentation difficulties outside eukaryotic cells, inefficient transfer of chromosomes to target cells, and variable mitotic stability. The quantity and quality of PSC-derived or reversibly immortalized stem/precursor cells that can transplant specific tissues are also critical determinants in the effectiveness of HAC-based tissue replacement therapies. Translating the HAC-based gene therapy platform remains difficult, but ongoing animal model research will move the HAC platform closer to clinical trials.

Genomic Context Vectors and Artificial Chromosomes for Cystic Fibrosis Gene Therapy

2007 ◽  
Vol 7 (3) ◽  
pp. 175-187 ◽  
Author(s):  
Massimo Conese ◽  
A. Christopher Boyd ◽  
Sante Di Gioia ◽  
Cristina Auriche ◽  
Fiorentina Ascenzioni
Keyword(s):  
Cystic Fibrosis ◽  
Gene Therapy ◽  
Cystic Fibrosis Gene ◽  
Genomic Context ◽  
Artificial Chromosomes

GENOMICS AND GENE THERAPY: Artificial Chromosomes Coming to Life

Science ◽  
2000 ◽  
Vol 290 (5495) ◽  
pp. 1308-1309 ◽  
Author(s):  
H. F. Willard
Keyword(s):  
Gene Therapy ◽  
Artificial Chromosomes

scholarly journals Transposon-Assisted Cloning and Traceless Mutagenesis of Adenoviruses: Development of a Novel Vector Based on SpeciesD

2006 ◽  
Vol 80 (16) ◽  
pp. 8100-8113 ◽  
Author(s):  
Zsolt Ruzsics ◽  
Markus Wagner ◽  
Andrea Osterlehner ◽  
Jonathan Cook ◽  
Ulrich Koszinowski ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Point Mutations ◽  
Cell Types ◽  
Sequence Information ◽  
Bacterial Artificial Chromosomes ◽  
Artificial Chromosomes ◽  
Coxsackievirus And Adenovirus Receptor ◽  
Species Specific ◽  
Adenovirus Receptor ◽  
Limited Sequence

ABSTRACT Until recently, adenovirus (Ad)-mediated gene therapy was almost exclusively based on human Ad type 5 (Ad5). Preexisting immunity and the limited, coxsackievirus and adenovirus receptor-dependent tropism of Ad5 stimulated attempts to exploit the natural diversity in tropism of the other 50 known human Ad serotypes. Aiming in particular at immunotherapy and vaccination, we have screened representative serotypes from different Ad species for their ability to infect dendritic cells. Ad19a, an Ad from species D, was selected for development as a new vector for vaccination and cancer gene therapy. To clone and manipulate its genome, we have developed a novel methodology, coined “exposon mutagenesis,” that allows the rapid and precise introduction of virtually any genetic alteration (deletions, point mutations, or insertions) into recombinant Ad bacterial artificial chromosomes. The versatility of the system was exemplified by deleting the E3 region of Ad19a, by specifically knocking out expression of a species-specific E3 gene, E3/49K, and by reinserting E3/49K into an E3 null Ad19a mutant. The technology requires only limited sequence information and is applicable to other Ad species. Therefore, it should be extremely valuable for the analysis of gene functions from any Ad species. In addition, a basic, replication-defective E1- and E3-deleted Ad19a vector expressing GFP (Ad19aGFP) was generated. This new vector based on species D Ads exhibits a very promising tropism for lymphoid and muscle cells and shows great potential as an alternative vector for transduction of cell types that are resistant to or only poorly transduced by conventional Ad5-based vectors.

scholarly journals Prospects for the Use of Artificial Chromosomes and Minichromosome-Like Episomes in Gene Therapy

2010 ◽  
Vol 2010 ◽  
pp. 1-16 ◽  
Author(s):  
Sara Pérez-Luz ◽  
Javier Díaz-Nido
Keyword(s):  
Gene Therapy ◽  
Genetic Diseases ◽  
Friedreich’S Ataxia ◽  
Friedreich's Ataxia ◽  
Regulatory Sequences ◽  
Mitotic Chromosomes ◽  
Physiological Regulation ◽  
Artificial Chromosomes ◽  
Recessive Mutations ◽  
Dividing Cells

Artificial chromosomes and minichromosome-like episomes are large DNA molecules capable of containing whole genomic loci, and be maintained as nonintegrating, replicating molecules in proliferating human somatic cells. Authentic human artificial chromosomes are very difficult to engineer because of the difficulties associated with centromere structure, so they are not widely used for gene-therapy applications. However, OriP/EBNA1-based episomes, which they lack true centromeres, can be maintained stably in dividing cells as they bind to mitotic chromosomes and segregate into daughter cells. These episomes are more easily engineered than true human artificial chromosomes and can carry entire genes along with all their regulatory sequences. Thus, these constructs may facilitate the long-term persistence and physiological regulation of the expression of therapeutic genes, which is crucial for some gene therapy applications. In particular, they are promising vectors for gene therapy in inherited diseases that are caused by recessive mutations, for example haemophilia A and Friedreich's ataxia. Interestingly, the episome carrying the frataxin gene (deficient in Friedreich's ataxia) has been demonstrated to rescue the susceptibility to oxidative stress which is typical of fibroblasts from Friedreich's ataxia patients. This provides evidence of their potential to treat genetic diseases linked to recessive mutations through gene therapy.

scholarly journals A new generation of human artificial chromosomes for functional genomics and gene therapy

2012 ◽  
Vol 70 (7) ◽  
pp. 1135-1148 ◽  
Author(s):  
Natalay Kouprina ◽  
William C. Earnshaw ◽  
Hiroshi Masumoto ◽  
Vladimir Larionov
Keyword(s):  
Gene Therapy ◽  
Functional Genomics ◽  
Artificial Chromosomes ◽  
New Generation

Mammalian artificial chromosomes — vectors for somatic gene therapy

1997 ◽  
Vol 118 (2) ◽  
pp. 135-142 ◽  
Author(s):  
F. Ascenzioni ◽  
P. Donini ◽  
H.J. Lipps
Keyword(s):  
Gene Therapy ◽  
Somatic Gene Therapy ◽  
Artificial Chromosomes ◽  
Somatic Gene
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