The CRISPR toolkit for Actinomyces includes both classic CRISPR-Cas9 related and CRISPR base editing based genetic manipulation tools for Actinomyces.
All plasmids are avaliable at Addgene. Furthermore, the toolkit is complemented by the sgRNA design software CRISPy-web.
pCRISPR-Cas9 (Addgene: 125686) can be used to generate deletion libraries by a single transformation; or to generate seamless deletion/insertion by providing editing templates.
pCRISPR-Cas9-ScaligD (Addgene: 125688) can be used to introduce small InDels.
pCRISPR-dCas9 (Addgene: 125687), CRISPRi, can be used to modulate transcriptional activity of target genes.
pCRISPR-cBEST (Addgene: 125689) and pCRISPR-aBEST (Addgene: 131464) can be used to convert C:G basepairs to T:A basepairs and A:T basepairs to G:C basepairs, respectively.
Figure: The CRISPR toolkit for Actinomyces.
References:
1. Tong, Y., Charusanti, P., Zhang, L., Weber, T. & Lee, S. Y. ACS Synth Biol 4, 1020-1029, (2015). doi:10.1021/acssynbio.5b00038
2. Tong, Y. et al. Proc Natl Acad Sci U S A 116, 20366-20375 (2019). doi:10.1073/pnas.1913493116
Contact: Prof. Tilmann Weber or Dr. Yaojun Tong