Difference between revisions of "Using CRISPRi to increase ethanol yield in E. coli MG1655"
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| − | {{Template: | + | {{Template:LaboratoryExercisesbyNLL}} |
==Overview== | ==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== | ==Laboratory exercises== | ||
| − | Day 1: | + | Day 1: <br> |
| − | Day 2: | + | Day 2: <br> |
| − | Day 3: | + | Day 3: <br> |
| − | Day 4: | + | Day 4: <br> |
| − | Day 5: | + | Day 5: <br> |
==Related modules== | ==Related modules== | ||
| − | [[ |In silico cloning of pdCas9 construct]] | + | #[[In silico cloning of pdCas9 construct |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 | ||
Latest revision as of 14:47, 2 August 2020
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
Day 1:
Day 2:
Day 3:
Day 4:
Day 5:
Related modules
- 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
