Using CRISPRi to increase ethanol yield in E. coli MG1655

CRISPRi as a teaching tool for inquiry-based investigations

<center>developed by Noreen L. Lyell
Instructor in the Department of Biological Engineering, MIT

Overview

Day 1:
Day 2:
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Day 4:
Day 5:

[[ |In silico cloning of pdCas9 construct]]
- Students perform steps used in classic cloning using free, online DNA manipulation interface using protocols 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 using protocols that can be completed in remote teaching format