Gene Inactivation and Tissue‐specific Metabolism

Matthew T Flowers, University of Wisconsin‐Madison, Madison, Wisconsin, USA
James M Ntambi, University of Wisconsin‐Madison, Madison, Wisconsin, USA

Published online: May 2008

DOI: 10.1002/9780470015902.a0021020

The application of gene inactivation strategies targeting genes with various metabolic functions has greatly advanced our understanding of metabolism at both the cellular and physiological level. Whole-body genomic ablation of a gene of interest may be achieved using transgenic methods to introduce a null allele. Alternatively, the Cre-loxP recombinase system allows for the generation of animals harbouring a tissue-specific gene inactivation. Another approach is to alter the messenger ribonucleic acid (mRNA) abundance of the target gene using RNA interference (RNAi) or antisense oligonucleotide (ASO) methods. Together, these strategies help elucidate gene function through association of metabolic phenotypes with gene inactivation at a defined genetic locus.

Keywords: transgenic mice; metabolism; gene knockout; RNA interference; Cre-loxP

Figure 1. Methods of gene inactivation: There are three main methods used to inactivate genes. (a) A whole-body gene knockout may be generated using transgenic animal technology. In this example, exon 3 of a gene of interest is being removed and replaced by homologous recombination with the neomycin resistance gene (NEOr), which is a selection marker. Regions of homology between the gene of interest and targeting vector are indicated by black rectangles. (b) A conditional knockout strategy allows for tissue-specific deletion of a gene. First, the gene of interest is altered with transgenic methods to introduce recombinase recognition sites (loxP or FRT) surrounding the region to be deleted. In the presence of the Cre or Flp recombinase, often under control of a tissue-specific promoter, the region between the recombinase recognition sites (indicated by black triangles) is deleted in cells expressing the recombinase. (c) RNAi or antisense oligonucleotide (ASO) methods utilize short sequences of nucleic acids complimentary to a target mRNA to induce mRNA destruction. In RNAi, the RNA-induced silencing complex (RISC) complex bound to the small interfering RNA (siRNA) recognizes and cleaves the complimentary mRNA sequence. ASOs form a DNA–RNA hybrid to block ribosomal function and promote cleavage of mRNA by ribonuclease (RNase) H.
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Related Sub-Topics:

Cell Biology of Cancer | Techniques in Cell Biology | Techniques and Tools in Molecular Biology | Biochemical and Biophysical Techniques | Nucleic Acids: Structure, Function, Metabolism
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Article Title: Gene Inactivation and Tissue‐specific Metabolism
 
How to Cite close
Flowers, Matthew T, and Ntambi, James M(May 2008) Gene Inactivation and Tissue‐specific Metabolism. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0021020]