The great majority of bacteria lack genetic tools which enable gene to associate with phenotypes systematically. CRISPR interference (CRISPRi) technology as a programmable method of reducing gene expression using a catalytically inactive Cas9 protein (dCas9) and a custom-designed guide RNA, has emerged as a versatile genetic tool to dissect the functions of essential and non-essential genes. In addition, CRISPRi libraries provides a valuable tool that target specific sets of genes or the entire genome straightforward. Consequently, genetic screens using CRISPRi libraries have contributed new insights into fundamental biology and molecular medicine including identifying functions for uncharacterized essential genes and drug modes of action.1
Microbiosci is committed to providing CRISPRi system service which enable gene to associate with phenotypes systematically. Our talented scientists have extensive experience in gene knockdown. They would research closely with you to offer help.
CRISPRi Library Design
Microbiosci provides a "one-stop shop" tailored to your gene research needs. An sgRNA library targeting genes of interest is synthesized. Oligonucleotides are amplified and cloned into expression plasmids, transformed into host strain expressing dCas9 protein, resulting in cell libraries. It's entirely customizable and flexible to evaluate the extent to which this gene is associated with relevant phenotypes.
Fig.1 A CRISPRi knockdown library in Streptococcus pneumoniae
The CRISPRi system has been used in the various bacterial cells, such as Escherichia coli, Bacillus, Clostridium, Corynebacterium, Lactobacillus, Mycobacterium, Pseudomonas, Staphylococcus, and Streptomyces.
Fig.2 CRISPRi provides robust repression in Bacillus subtilis
Fig.3 CRISPRi provides robust repression in P. aeruginosa
Microbiosci is the premier institution to provide professional and comprehensive genome editing services in microbial. We have years of experience to meet your specific project needs in using the CRISPR technology to add value to your research project.
1. Peters, J. M., Koo, B., Patino, R., Heussler, G. E., Hearne, C. C., Qu, J., ... & Rosenberg, O. S. (2019). Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nature microbiology, 4(2), 244-250.
2. Tong, Y., Weber, T., & Lee, S. Y. (2019). CRISPR/Cas-based genome engineering in natural product discovery. Natural Product Reports,.
3. Liu, X., Gallay, C., Kjos, M., Domenech, A., Slager, J., Van Kessel, S. P., ... & Veening, J. (2017). High‐throughput CRISPRi phenotyping identifies new essential genes in Streptococcus pneumoniae. Molecular Systems Biology, 13(5), 1-18.
4. Peters, J. M., Colavin, A., Shi, H., Czarny, T. L., Larson, M. H., Wong, S., ... & Gross, C. A. (2016). A Comprehensive, CRISPR-based Functional Analysis of Essential Genes in Bacteria. Cell, 165(6), 1493-1506.
5. Tan, S. Z., Reisch, C. R., & Prather, K. L. (2018). A Robust CRISPR Interference Gene Repression System in Pseudomonas. Journal of Bacteriology, 200(7).