Novel Method to Load Multiple Genes onto a Mammalian Artificial Chromosome
| Autor: | Robert L. Katona, Tünde Praznovszky, Vilmos Tubak, Andor Udvardy, Anna Tóth, Gyula Hadlaczky, Katalin Fodor |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Heredity
Mutant Gene Expression Biochemistry Polymerase Chain Reaction Mice Nucleic Acids Cricetinae Molecular Cell Biology Induced pluripotent stem cell In Situ Hybridization Fluorescence Genetics Multidisciplinary Chromosome Biology Immunochemistry Gene targeting Genomics Gene Therapy Enzymes Medicine Genetic Engineering Adult stem cell Research Article Biotechnology Chromosome Structure and Function Histology Science Genetic Vectors Chromosomes Artificial Mammalian Computational biology CHO Cells Biology Marker gene Molecular Genetics Cytogenetics Model Organisms Cricetulus Genomic Medicine Animals Gene Selectable marker DNA Primers Tissue Engineering Base Sequence Proteins Embryonic stem cell Disease Models Animal Genes Bionanotechnology |
| Zdroj: | PLoS ONE PLoS ONE, Vol 9, Iss 1, p e85565 (2014) |
| ISSN: | 1932-6203 |
| Popis: | Mammalian artificial chromosomes are natural chromosome-based vectors that may carry a vast amount of genetic material in terms of both size and number. They are reasonably stable and segregate well in both mitosis and meiosis. A platform artificial chromosome expression system (ACEs) was earlier described with multiple loading sites for a modified lambda-integrase enzyme. It has been shown that this ACEs is suitable for high-level industrial protein production and the treatment of a mouse model for a devastating human disorder, Krabbe’s disease. ACEs-treated mutant mice carrying a therapeutic gene lived more than four times longer than untreated counterparts. This novel gene therapy method is called combined mammalian artificial chromosome-stem cell therapy. At present, this method suffers from the limitation that a new selection marker gene should be present for each therapeutic gene loaded onto the ACEs. Complex diseases require the cooperative action of several genes for treatment, but only a limited number of selection marker genes are available and there is also a risk of serious side-effects caused by the unwanted expression of these marker genes in mammalian cells, organs and organisms. We describe here a novel method to load multiple genes onto the ACEs by using only two selectable marker genes. These markers may be removed from the ACEs before therapeutic application. This novel technology could revolutionize gene therapeutic applications targeting the treatment of complex disorders and cancers. It could also speed up cell therapy by allowing researchers to engineer a chromosome with a predetermined set of genetic factors to differentiate adult stem cells, embryonic stem cells and induced pluripotent stem (iPS) cells into cell types of therapeutic value. It is also a suitable tool for the investigation of complex biochemical pathways in basic science by producing an ACEs with several genes from a signal transduction pathway of interest. |
| Databáze: | OpenAIRE |
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