All design parameters are automatically tracked, making it very easy to reproduce & compare experimental outcomes. All workflows further come with html protocols, so it's super easy to just get started in the lab.

6/8We then demonstrated how the high efficiencies of CASCADE-Cas3 can be used for streamlined host construction by substitution of large genomic regions with PhiC31 integration sites - allowing multicopy expression of BGCs. Reintroduction of the act BGC resulted in significantly higher production.

4/8 Furthermore, efficiencies appeared to be less dependant on which spacer was used, indicating less background/off-target activity. What about other streptomycetes? We picked S. albidoflavus, S. venezuelae, as well as NBC1270, an isolate from our collection, for further experiments.

3/8 So we were wondering - is CASCADE-Cas3 much better suited for genome engineering of streptomycetes? Targeting the act BGC of S. coelicolor, we got some very strong initial data supporting this hypothesis. We obtained very precise deletions with robustly high efficiencies.

2/8 The minimal type I-C CASCADE-Cas3 is fascinating. A complex consisting of Cas5, Cas7, and Cas8 binds the target site and recruits the processive nuclease Cas3, which generates recombinogenic overhangs! Sounds great for homologous recombination!