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CRISPR-Cas applied to TGF-beta induced EMT Virtual Lab

Learn how to detect the hallmarks of EMT using immunofluorescence and knock out the correct gene to revert the process using CRISPR

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CRISPR-Cas applied to TGF-beta induced EMT

About this Simulation

CRISPR may sound like a brand of cereals, but it certainly isn’t! In this simulation, you will learn to identify the molecular hallmarks of the Epithelial-to-Mesenchymal Transition (EMT) molecular process in breast cancer cells. You will induce EMT by adding TGF-beta to the cells, and, after identifying the hallmarks with immunofluorescence labeling, you will use the cutting-edge CRISPR-Cas9 technology to try to revert this process, which is one of the main causes of cancer malignancy.

Set up your own immunofluorescence protocol

In your first mission, you will be able to perform an immunostaining protocol by making decisions on the key steps of the process. Each decision can get you closer to the correct outcome, so think carefully and choose wisely!

Use CRISPR to knock out a specific gene

Once you have learned what the molecular hallmarks of EMT are, you will be able to dive into a eukaryotic cell to learn how the CRISPR-Cas9 technique works at a molecular level. Then, you will learn to design one of the CRISPR-Cas9 components by solving a fun puzzle. Last, you will select the target gene to knock out and your virtual lab assistant, Dr. One, will help you with the last part of the immunostaining process so you can directly evaluate your results.

Repeat your experiment as many times as you want

At the end of each mission, you will be able to repeat the experiment if the outcome was not the desired one, giving you the possibility of learning from your mistakes. Luckily, Dr. One will be there to help you with some of the tasks. In the end, you will be able to complete an experiment that takes weeks in just less than one hour!

Will you be able to set the right protocols in place, solving any issues that arise, to understand the molecular changes linked to the TGF-beta induced EMT and evaluate the possibility of using CRISPR-Cas9 as a possible therapeutic approach in breast cancer?

Learning Objectives

At the end of this simulation, you will be able to:

  • Set up a protocol to get answers to a biological question related to the EMT process

  • Acknowledge mistakes in the protocols and understand how to critically overcome them

  • Properly prepare cells for the immunostaining procedure according to the localization of the target protein

  • Select a suitable secondary antibody for an immunofluorescence experiment

  • Analyze immunofluorescence results

  • Describe the basics of CRISPR-Cas technique

  • Design a guide RNA construct for knock-out strategies

  • Evaluate CRISPR-Cas9 results

Techniques in Lab

At the end of this simulation, you will be able to:

  • CRISPR-Cas9
  • Immunofluorescence

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