In this module you will use a modified Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system to modulate transcription in E. coli. Specifically, you will use a CRISPR-based interference (CRISPRi) system to target a gene within the fermentation pathway of E. coli such that either ethanol or acetate production is increased.
- In silico cloning of pdCas9 construct
- Students perform steps used in classic cloning using free, online DNA manipulation interface with laboratory exercises that can be completed in remote teaching format
- Useful when timing doesn't allow for at-the-bench cloning laboratory exercise
- Provides background details on methods used for plasmid construction that are important in laboratory exercises focused on the steps after cloning
- Steps for optional at-the-bench confirmation digest laboratory exercise included
- Optimizing sgRNA design to further increase ethanol yield (coming soon!)
- Students analyze datasets collected in previous semesters to design an optimized sgRNA sequence with laboratory exercises that can be completed in remote teaching format