Using CRISPRi to increase ethanol yield in E. coli MG1655

Overview

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.

Laboratory exercises

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Related modules

1. 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
2. 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