Multiple strategies for gene transfer, expression, knockdown, and chromatin influence in mammalian cell lines and transgenic animals |
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Authors: | Félix Recillas-Targa |
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Institution: | (1) Instituto de Fisiologia Celular, Departamento de Genética Molecular, Universidad Nacional Autonoma de México, Apartado Postal 70-242, 04510 México, D.F. |
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Abstract: | Manipulation of the eukaryotic genome has contributed to the progress in our knowledge of multicellular organisms but has
also ameliorated our experimental strategies. Biological questions can now be addressed with more efficiency and reproducibility.
There are new and varied strategies for gene transfer and sequence manipulation with improved methodologies that facilitate
the acquisition of results. Cellular systems and transgenic animals have demonstrated their invaluable benefits. In this review,
I present an overview of the methods of gene transfer with particular attention to cultured cell lines and large-scale sequence
vectors, like artificial chromosomes, with the possibility of their manipulation based on homologous recombination strategies.
Alternative strategies of gene transfer, including retroviral vectors, are also described and the applications of such methods
are discussed. Finally, several comments are made about the influence of chromatin structure on gene expression. Recent experimental
data have shown that for convenient stable transgene expression, the influence of chromatin structure should be seriously
taken into account. Novel chromatin regulatory and structural elements are proposed as an alternative for proper and sustained
gene expression. These chromatin elements are facing a new era in transgenesis and we are probably beginning a new generation
of gene and cancer therapy vectors. |
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Keywords: | Gene transfer gene expression antisense transient transfection stable expression retroviral infection gene therapy microcell fusion position effects chromatin Locus Control Region insulator artificial chromosomes recombination systems viral vector |
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