Learning Lab: Knockout! A CRISPR/Cas Gene Targeting Lab

This lab is easy for teachers to prepare, makes gene targeting accessible to students, and packs a powerful punch!Become a genetic engineer and use CRISPR/Cas to disable a gene in bacteria. Scientists use the CRISPR/Cas system to alter the DNA in living cells. Now you can bring this cutting-edge technique to your advanced biology class! In this lab, E. coli cells which express the lacZ gene from a plasmid will be transformed with another plasmid containing the cas9 gene and a custom guide RNA. This guide RNA contains a sequence complementary to the lacZ gene so, in transformed cells, the guide RNA will target Cas9 to cut the lacZ gene, disabling it. Students can easily identify their CRISPR/Cas knockouts using the color of the bacterial colonies. Cells which express lacZ are able to break down the chemical X-gal into a blue product. Cells with a functional lacz will be blue, while cells in which the CRISPR/Cas system has successfully disabled lacZ will revert to a white phenotype.

Note: This is a technically advanced lab that requires a 2-hour class period. Students should have prior experience with bacterial transformation.

Techniques: Micropipetting, bacterial transformation, gene targeting
Topics: Genome editing, biotechnology, molecular biology
Time required: Two 45-min or one 90-min class period
Level: Advanced high school through college
For educational use only.

Reagents for eight lab groups

Ship-out date: 21-working day
Brand Link: https://www.minipcr.com/product/knockout/