Crispr Experiments at Your House?

This week, I managed to get my hands on a Crispr Kit, that would essentially allow me to program bacteria. Now I can’t tell you how excited I was for this! Not only was I able to pexperiment but I was able to learn so much from it. And I would just like to share my experiences and what I learned over the journey!

Transforming My Home Into A Lab

Right as I opened up the Crispr Kit, I found a plethora of items. Normal equipment, but also the bacteria and Crispr components! Nothing was missing, and I found myself eager to experiment.

Preparing the Bacteria

The first step in this bacteria growing endeavor is first to give them a place to grow! Included with the kit were 2 agar media(powders) that would mix and turn into agar, which would essentially feed the bacteria and allow for them to proliferate.

Here is a picture of the LB Agar being fully mixed. I then poured this liquid into six Petri dishes and let them cool!

After I let the agar cool down, I went and moved on to the next step. However, since my pouring technique was not on par, I only used 3 Petri dishes to grow the bacteria, as the other dishes were not fully covered with the agar or were just pretty bumpy. It isn’t as easy as it looks folks!

The second stage was actually to prepare the bacteria. The bacteria was an E. Coli that was non-pathogenic(not dangerous) and it arrived freeze-dried. This meant that the bacteria was dormant, and I had to wake it up!

To do that, I added water into the capsule that the bacteria was stored in, and then rehydrated the bacteria, allowing it to live again! This mixture would then be swabbed over the Petri dishes and I allowed them to grow for a few days!

Finally Editing the Bacteria

I think that the coolest part of the experiment was using the pipette. It was amazing to see such precision when it came to measuring, and it makes you feel like a real scientist! Alright, back to the experiment.

After letting the bacteria grow for a few days, I was finally able to begin the Crispr process. I first added a transformation mix, which would make the bacteria cocompetent. This meant that the bacteria would be more accepting of foreign particles(and our DNA and Crispr) as the cell wall would be weakened.

Adding the Crispr Components

Here you see me holding three tubes holding the Cas 9 proteins, template RNA, and the guide RNA. These three components make up the whole Crispr system!

I scooped up parts of the bacteria on the plate and mixed them in a tube with the Crispr components, and I let them incubate for thirty minutes in the fridge. I then quickly incubated them in hot water and added a mixture of LB media. LB stands for lysogeny broth and it is also used in the agar. This stew helps to feed the bacteria and allows it to grow. But more importantly, it allows for the bacteria to incorporate the foreign DNA and edit itself with Crispr. After adding some LB media, I let the mixture incubate overnight at room temperature.

Plating the New Bacteria!

After I let the bacteria incubate, I made 7 new plates full of LB Strep/Khan. These plates are different from the first ones, as they essentially won’t allow the original bacteria to grow. However, our experiment will edit the bacteria so that it will be able to grow just fine on this deadly material.

Like before, I streaked these plates with the fully incubated bacteria, and I let them sit for a day or so. And when I came back I was surprised! Although most of the plates produced molds, one plate managed to have exactly what I was looking for!

I let this batch grow a bit more before I took the picture, explaining why there is so much growth.

Compared to the picture in the protocol, it was practically spot on, except for the fact that I streaked it a bit more and left it out for longer. This was one out of seven plates. Instead of doing separate experiments, I did them all at once, so I had huge batches. This meant that I had a lot of materials to work with, and thus helped me to save time and find success.

What I learned

Before doing this experiment, I knew Crispr, but I didn’t know it enough. I learned a lot from going through this experiment and I learned some cool things like pipetting techniques. Although I still have so much to learn when it comes to Crispr and just gene editing in general, I hope that I will be able to learn more and more, and maybe use this knowledge for good one day!

Takeaways:

• You can experiment with Crispr for less than 200 dollars
• Your home can become a lab to experiment in!
• Crispr is so complex yet amazing