The Next Super Soldier With The Help of CRISPR

8 min readDec 30, 2021

Steve Rogers who started out being a scrawny boy turned into a super-soldier we now know as Captain America. We all probably dreamt about what if we turned into a super-soldier and could just get magically muscular without having to go to the gym.

But what if I told you it was possible, that we could manufacturer our own super-soldier serum with an emerging technology called CRISPR. CRISPR-Cas9 to put it simply is a protein that can edit our DNA. Having so many interesting applications from cures to genetic diseases to super strength. So let’s walk through how we could potentially pull it off. First, we need to talk about Benchling and how it was the perfect software for the job.

What is Benchling? 💻

Benchling was founded on January 1, 2012. But the idea of this company started in an MIT dorm which at the beginning was a side project for Sajith Wickramasekara and his classmate Ashu Singhal. But this started as a small side project turned into a billion-dollar company. They noticed most of the work done in life sciences was still being done on paper! This is crazy so Benchling came in to make it easier for scientists and students to collaborate and create innovations that will change the future forever.

One feature that we are going to take a closer look at is Benchling’s CRISPR tools. As one amazing perk of Benchling is that you can make a gRNA in the site itself. Giving us the choice of choosing our Protospacer Adjacent Motif (PAM) and Guide Length with a bunch of other options we will discuss later. The last main reason it’s an amazing software is that it gives you predictions for on and off-target effects of the Cas9’s binding site. Which is more useful than you might think.

The Basics of Editing DNA ✂️

Before we go on to making gene edits that can turn you super muscular we need to learn the basics of how Benchling works and how you actually use the CRISPR tools. So let’s begin…

Our Practice Run 🧬

Step 1: Import a DNA Sequence

Before we can do anything we actually need a DNA sequence to edit. Without it, we are basically editing air. For this practice example let’s choose a random DNA sequence like BRCA2. So to import the sequence into Benchling you can go over and press the briefcase icon that should show you a blue plus button. After pressing the plus button it will give you a slide down option where you can select “DNA Sequence” and then “New DNA Sequence”

There you can either import the DNA Sequence or use one cool feature of Benchling which is “Search External Database” and in the search bar you can type in the gene you want and it will find it for you! After you search up BRCA2 you just have to make sure it’s going to the right folder and press import.

Step 2: What your Seeing

Now you should have reached a page like mine where you can see the “Sequence Map” on the left side and the “Linear Map” on the right side. At first, this might seem like a lot, but to break it down it’s very simple. But before I break it down you need to know what Exons and Introns are.

Exons and Introns?

A good way to think of this is that exons are the ones coding for the proteins and the introns are just there as almost spacers. Introns still do have a purpose but for now, just think of them as doing nothing and the exons actually coding for the protein.

Sequence Map

On the left, you can see the Adenine (A), Thymine (T), Cytosine (C), and Guanine (G) of the Gene. With that, you should also see annotations that say Exon with some number behind it. The Exons are automatically put there and numbered to make it easier for you to understand while reading the gene. Lastly, you can also see the amino acids in different colours under the Exon section.

Linear Map

On the right side, you should see the linear display that shows the whole gene as a line with markers for exons and the lines that are shooting up from the gene are enzymes with their own complicated name. On that side, you should also see the base pair length which is really cool as you can see that in our example there is 40742. The base-pair length basically is the word count in an essay but instead of words, it’s base pairs.

Step 3: CRISPR Tool 🧫

Hopefully, you now understand a bit more about what you’re actually looking at. The next step is to set up the Guide RNA (gRNA) for the CRISPR-Cas9. The gRNA is what Cas9 is going to use to find the piece of the gene you want it to leave. So what you have to do is look at the right side of the screen and you should see 7 icons and the icon we need is the sixth one that has a target as it’s an icon. When you click on it you will see two options and you have to choose the blue button named “Design and Analyse Guides”.

After you do that you will get to a page where it says “Desgin Type”, “Guide Length”, “Genome”, and “PAM”. We are only going to look at the last three for what we need. The Guide Length is the length of the guide RNA. With the standard number being 20. Next is Genome this is where you type in what we selected in the beginning when importing the gene from its genome which for us was Rnor_6.0(rn6, Rattus Norvegicus). Lastly is PAM which stands for Protospacer Adjacent Motif. The PAM for those who don’t know is a small sequence of DNA that the Cas9 needs to make a cut. But the cool part is that there are different types of PAM like the most common NGG or it could be something different like NAG instead. For our example, we only need to NGG so the default. After that press finishes so we could get to the next step.

Step 4: Selecting Target Region 🎯

After we chose the parameters of our gRNA we need to actually now select our target region. The best way to do it is by highlighting the target region but you could also type it in. After you select a target region you press the green plus icon and you should get to the next stage. In this stage, you will get a choice of what guide sequence you want to use. With the On and Off-Target effects being scored 0–100 with getting a high score for both of them being really good. So once you chose the one you want to use just press assemble and for the last step you just need to choose an expression vector which will basically make it so that Cas9 can have a promoter. After pressing next you are done and you have successfully made a Plasmid.

Now that we have gone and made a plasmid how can that help give someone super strength?

My Project 💪

The Gene MSTN, this gene codes for something called Myostatin. This protein is found in muscle tissue. Its primary function is to negatively regulate muscle growth. As to much muscle mass can cause damage to your bones so as a way to prevent that from happening our body regulates our muscle growth with Myostatin. But here is where it gets interesting as mutations are a natural occurrence and when there is a mutation in Myostatin that makes it so the body can’t produce enough of the Myostatin protein. This can result in people gaining large amounts of muscles mass with less effort.

The mutation’s name is a special kind of Muscle Hypertrophy or the “Hercules Gene”. With some real-world examples being Eddie Hall, the winner of the 2017 World’s Strongest Man Competition had this gene. Allowing him to gain more muscle mass than the average man! Another example of it actually being done is when they were able to genetically modify sheep to have the same or at least similar mutation.

Simulating MSTN 🏋️

So now that we have learned about Benchling we can use the same steps above to edit the gene of MSTN. The specific binding area for this project was exon 1 as I think it’s the best and most accurate place the mutation would be after going through resources on Muscular Hypertrophy (Hercules Gene). So that is what I did, I imported the gene into Benchling. Started to make my gRNA and chose a binding site that would have the most on target and less off-target effects.

With my final product, I got a plasmid with Cas9 that inside of it has the potential to turn you into the next super-soldier.

The Future ✨

CRISPR-Cas9 has accelerated the process of gene editing and has reshown the world the horizon of possibilities gene therapies have to offer. As I was able to make a plasmid that could have the potential to turn someone into the next Captain America. Imagine the countless more groundbreaking innovations could we do. As the word, science fiction is slowly turning into our new reality.