20.109(F19):Module 2

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20.109(F19): Laboratory Fundamentals of Biological Engineering

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Fall 2019 schedule        FYI        Assignments        Homework        Class data        Communication
       1. Measuring genomic instability        2. Modulating metabolism        3. Testing chemical probes              

Module 2

Lecturer: Noreen Lyell
Instructors: Noreen Lyell, Leslie McClain and Becky Meyer
TAs: Colin Kim and Shelbi Parker
Lab manager: Hsinhwa Lee


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 system to target a gene within the fermentation pathway of E. coli such that either ethanol or acetate production is increased.

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Lab links: day by day

M2D1: Complete in silico cloning of pdCas9
M2D2: Design gRNA for CRISPRi
M2D3: Generate gRNA plasmid
M2D4: Journal club I
M2D5: Confirm gRNA sequence in pgRNA
M2D6: Journal club II
M2D7: Induce CRISPRi system
M2D8: Measure fermentation products


Research article
Journal club presentation


CRISPR interference (CRISPRi) for sequence-specific control of gene expression., Nature Methods. (2013) 8: 2180-2196.

Notes for teaching faculty

Prep notes for M2