Design of vectors for transgene expression: The use of genomic comparative approaches.
The design of transgenes has always been limited by the extent of available information on the endogenous locus whose expression pattern had to be replicated. Those genes whose expression domain had not been entirely documented resulted, usually, in transgenes with an unpredictable expression patterns and suboptimal performance in transgenic animals. The use of genomic comparative approaches, highlighting evolutionary conserved homologous DNA sequences, helps to identify crucial regulatory elements that are associated to a given expression domain. The inclusion of these conserved regulatory sequences in transgenic constructs would normally result in optimal expression levels of transgenes in recipient animals. The use of artificial chromosome-type transgenes usually ensures the inclusion of these preserved regulatory elements that are required for the faithful expression of the gene. These constructs could also contain insulators, a subset of regulatory sequences whose application is being addressed in transgenesis. Therefore, the generation of transgenic animals with genomic-type constructs is the recommended approach to achieve optimal transgene expression, according to the expected pattern of the corresponding endogenous locus.
Montoliu
Lluís
L
Department of Molecular and Cellular Biology, Centro Nacional de Biotecnologia (CNB-CSIC), Campus de Cantoblanco, C/ Darwin 3, 28049 Madrid, Spain. montoliu@cnb.uam.es
Roy
Rosa
R
Regales
Lucía
L
García-Díaz
Angel
A
D000818:Animals; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D023281:Genomics; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D008894:Milk Proteins / Q000235:genetics*; D014442:Monophenol Monooxygenase / Q000235:genetics*; D050436:Regulatory Elements, Transcriptional / Q000235:genetics*; D019076:Transgenes / Q000235:genetics*
D008894:Milk Proteins; C024471:whey acidic proteins; D014442:Monophenol Monooxygenase