Rath Devashish, Amlinger Lina, Hoekzema Mirthe, Devulapally Praneeth Reddy, Lundgren Magnus
Department of Cell and Molecular Biology, Uppsala University, SE-751 24 Uppsala, Sweden.
Department of Cell and Molecular Biology, Uppsala University, SE-751 24 Uppsala, Sweden
Nucleic Acids Res. 2015 Jan;43(1):237-46. doi: 10.1093/nar/gku1257. Epub 2014 Nov 30.
Methods that permit controlled changes in the expression of genes are important tools for biological and medical research, and for biotechnological applications. Conventional methods are directed at individually changing each gene, its regulatory elements or its mRNA's translation rate. We demonstrate that the CRISPR-associated DNA-binding Cascade complex can be used for efficient, long-lasting and programmable gene silencing. When Cascade is targeted to a promoter sequence the transcription of the downstream gene is inhibited, resulting in dramatically reduced expression. The specificity of Cascade binding is provided by the integral crRNA component, which is easily designed to target virtually any stretch of DNA. Cascade targeted to the ORF sequence of the gene can also silence expression, albeit at lower efficiency. The system can be used to silence plasmid and chromosome targets, simultaneously target several genes and is active in different bacterial species and strains. The findings described here are an addition to the expanding range of CRISPR-based technologies and may be adapted to additional organisms and cell systems.
能够控制基因表达变化的方法是生物学和医学研究以及生物技术应用的重要工具。传统方法旨在单独改变每个基因、其调控元件或其mRNA的翻译速率。我们证明，与CRISPR相关的DNA结合级联复合物可用于高效、持久且可编程的基因沉默。当级联复合物靶向启动子序列时，下游基因的转录受到抑制，导致表达显著降低。级联结合的特异性由完整的crRNA组分提供，该组分很容易设计成靶向几乎任何一段DNA。靶向基因ORF序列的级联复合物也能使表达沉默，尽管效率较低。该系统可用于使质粒和染色体靶点沉默、同时靶向多个基因，并且在不同的细菌物种和菌株中均有活性。此处描述的研究结果是对基于CRISPR的技术不断扩展的范围的补充，并且可能适用于其他生物体和细胞系统。
