Alessandra Stefan, Alessandro Tonelli, Flavio Schwarz, Alejandro Hochkoeppler
Department of Industrial Chemistry, University of Bologna, Viale Risorgimento 4, 40136 Bologna, CSGI, University of Florence, Italy.
BMB Rep. 2008 Aug 31;41(8):568-74. doi: 10.5483/bmbrep.2008.41.8.568.
Antisense RNA molecules are powerful tools for controlling the expression of specific genes but their use in prokaryotes has been limited by their unpredictable antisense effectiveness. Moreover, appreciation of the molecular mechanisms associated with silencing in bacteria is still restricted. Here we report our attempts to define an effective antisense strategy in E. coli, and to dissect the observed silencing process. Antisense constructs complementary to different regions of lacZ were investigated, and silencing was observed exclusively upon expression of antisense RNA hybridising the 5'UTR of lac messenger. The level of lacZ mRNA was reduced upon expression of this antisense construct, and the silencing competence was found to be closely associated with its stability. These observations may help in the design of antisense molecules directed against prokaryotic genes.
反义RNA分子是控制特定基因表达的有力工具，但它们在原核生物中的应用受到其不可预测的反义效果的限制。此外，对细菌中与沉默相关的分子机制的认识仍然有限。在此，我们报告了我们在大肠杆菌中定义有效反义策略并剖析观察到的沉默过程的尝试。研究了与lacZ不同区域互补的反义构建体，并且仅在与lac信使的5'UTR杂交的反义RNA表达时观察到沉默。该反义构建体表达后，lacZ mRNA水平降低，并且发现沉默能力与其稳定性密切相关。这些观察结果可能有助于设计针对原核基因的反义分子。
