Metabolic engineering efforts often demand controlling the expression level of multiple genes. Furthermore, the ability to precisely regulate gene expression is significant to understand desired genes' functions. CRISPR-based gene activation, termed CRISPRa, utilizes dCas9 fusion protein to bind various transcription activators.1 dCas fusion protein provides a simple and robust technology for gene activation, and can precisely target DNA sequence with the aid of the sgRNA. The dCas9 complex guided by crRNA binds to the upstream promoter regions and recruits the RNA polymerase, and further activates transcription.2
Furthermore, together with existing tools for CRISPRi gene repression, these bacterial activators enable programmable control over multiple genes with simultaneous activation and repression. Consequently, bacterial activators will offer the basis for engineering synthetic bacterial devices with applications consist of diagnostics, therapeutics, and industrial biosynthesis.3
Fig.1 CRISPR activation in bacteria enables complex multi-gene expression programs.
Due to the simpler design, better performance and less sequence constraint, the ability of CRISPRa to activate gene expression is more powerful.2 For instance, fusion of the ω subunit of RNA polymerase to the dCas protein can upregulate gene expression up to 3-fold in E.coli.4
With dynamically-controlled CRISPRa systems, Creative Biogene is committed to build more complex synthetic bacterial devices with a wide range of applications in industrial biosynthesis. Our talented scientists would research closely with you to offer help. We have years of experience to meet your specific project needs in using the CRISPR technology to add value to your research project.
1. Dominguez, A. A. , Lim, W. A. , & Qi, L. S. . (2015). Beyond editing: repurposing crispr–cas9 for precision genome regulation and interrogation. Nature Reviews Molecular Cell Biology.
2. Yao, R., Liu, D., Jia, X., Zheng, Y., Liu, W., & Xiao, Y. (2018). CRISPR-Cas9/Cas12a biotechnology and application in bacteria. Synthetic and Systems Biotechnology, 3(3), 135–149.
3. Dong, C. , Fontana, J. , Patel, A. , Carothers, J. M. , & Zalatan, J. G. . (2018). Author correction: synthetic crispr-cas gene activators for transcriptional reprogramming in bacteria. Nature Communications, 9(1).
4. Albert, L. , & Lei, Q. . (2017). Genetic and epigenetic control of gene expression by crispr–cas systems. F1000Research, 6, 747-.
5. Tong, Y., Weber, T., & Lee, S. Y. (2019). CRISPR/Cas-based genome engineering in natural product discovery. Natural Product Reports,.