Difference between revisions of "Using CRISPRi to increase ethanol yield in E. coli MG1655"
From Course Wiki
Noreen Lyell (Talk | contribs) (→Related modules) |
Noreen Lyell (Talk | contribs) (→Overview) |
||
| (3 intermediate revisions by one user not shown) | |||
| Line 5: | Line 5: | ||
==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== | ||
| Line 16: | Line 18: | ||
==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 | + | #*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 | #*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 | #*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 | #*Steps for optional at-the-bench confirmation digest laboratory exercise included | ||
| − | |||
#Optimizing sgRNA design to further increase ethanol yield (coming soon!) | #Optimizing sgRNA design to further increase ethanol yield (coming soon!) | ||
| − | #*Students analyze datasets collected in previous semesters to design an optimized sgRNA sequence | + | #*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
