Montoliu Lluís, Roy Rosa, Regales Lucía, García-Díaz Angel
Department of Molecular and Cellular Biology, Centro Nacional de Biotecnologia (CNB-CSIC), Campus de Cantoblanco, C/ Darwin 3, 28049 Madrid, Spain.
Comp Immunol Microbiol Infect Dis. 2009 Mar;32(2):81-90. doi: 10.1016/j.cimid.2007.11.013. Epub 2008 Mar 6.
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.
转基因的设计一直受到内源性基因座可用信息范围的限制，而转基因的表达模式必须复制内源性基因座的表达模式。那些表达域未被完全记录的基因，通常会导致转基因在转基因动物中具有不可预测的表达模式和次优性能。使用基因组比较方法，突出进化保守的同源DNA序列，有助于识别与给定表达域相关的关键调控元件。将这些保守调控序列纳入转基因构建体通常会使转基因在受体动物中达到最佳表达水平。使用人工染色体型转基因通常可确保纳入基因忠实表达所需的这些保留调控元件。这些构建体还可包含绝缘子，这是一类调控序列，其在转基因中的应用正在研究中。因此，根据相应内源性基因座的预期模式，使用基因组型构建体生成转基因动物是实现最佳转基因表达的推荐方法。
