Biomedical Engineering Reference
In-Depth Information
Gene Expression
in Mammalian Cells
Felix Recillas-Targa, Georgina Guerrero, Martın Escamilla-del-Arenal
and Hector Rincon-Arano
Instituto de Fisiologıa Celular, Departamento de Genetica Molecular, Universidad Nacional Autonoma de
Mexico, Apartado, Mexico
7.1 Introduction
Understanding of the sophisticated processes of gene expression in mammalian cells neces-
sitates manipulation of the eukaryotic genome. Such manipulation has greatly contributed
to the progress in our knowledge of multicellular organisms, albeit having forced amelio-
ration of our experimental strategies. There are new and varied strategies for gene transfer
and sequence manipulation with improved methodologies that facilitate the acquisition of
results. Recent experimental data has shown that, for convenient stable transgene expression
analysis, the influence of chromatin structure should be seriously taken into consideration.
Novel regulatory and structural chromatin elements have been proposed as necessary for
appropriate and sustained gene expression. These chromatin elements are facing a new era
in transgenesis, and we are probably beginning to see a new generation of gene and cancer
therapy vectors. In this chapter, we first briefly discuss and describe several strategies for
gene transfer and genome manipulation, and then two alternative methods are presented for
the study of gene expression in mammalian cells. In the first method, particular attention
is paid to the genomic integration context of transgenes, while the second is designed to
maintain chromosomal integrity containing the genomic sequences under study.
Starting with recombinant DNA technology, a large spectrum of applications has emerged
in gene manipulation area. In particular, intense efforts have been made to transfer genetic
information to cell lines, primary cell cultures, diverse organisms and tissues, and in the
generation of genetically modified organisms. Without a doubt, gene transfer has been
critical in the advancement of our knowledge related to general phenomena such as gene
