What

A CRISPRi tool for knockdown based module development.

Blockdiagram

Partsdiagram

Why

Instead of complete removal, it may be useful to be able to knockdown the function of a part, device, circuit or module in a genome.

How

Clustered Regularly Interspaced Short Palindromic Repeats Inhibition (CRISPRi) -based system coupled with CRISPathBrick arrays.

What Is Success

A versatile platform for knockdown-based development.


Work In Progress

Histidine Knockdown

We constructed a protospacer to knockdown the histidine biosynthetic pathway by targeting the HisBHAFI operon. The CRISPRi construct was transformed into E. coli MG1655, which were grown in LB, minimal media, and minimal media supplemented with histidine. Optical density was measured over 36 hours. Points are mean over 12 replicates; error bars are standard error.

Histidine Knockdown

Sample Media KD
LB.NP LB  
LB.P LB Y
Aux.NP Minimal  
Aux.P Minimal Y
His.NP Minimal + His  
His.P Minimal + His Y
His2.NP Minimal + His  
His2.P Minimal + His Y