Controlling the KRas Oncogene With Crispr

Cancers cause millions of deaths every year. They are some of the deadliest diseases, and yet they are a product of our own human bodies, rather than some outside source.

What Causes Cancer?

Usually, this is perfectly fine, as our bodies immune system would be able to get rid of these bad cells. However, as time progresses, more and more of these annoying little cells manage to escape detection and our immune system weakens more and more. This is why the risk of cancer goes further up the older you get.

Oncogenes

Imagine a cell as a car and a proto-oncogene as the gas pedal. You can use it to accelerate, but you still have control. Now imagine that pedal gets stuck, and now you can’t stop accelerating. That is basically an oncogene!

Tumor Supressor Genes

The most common abnormality to a tumor suppressor gene would be to the TP53 gene. Almost half of all cancers have mutations in this gene, and a lot of research has been done to see how we can activate these genes again!

Going back to the car analogy, the tumor suppressor gene acts as brakes, to stop the car when needed. Now imagine if the car had brakes that didn’t work! Now, that is a surefire way to get into a car wreck, and a cancer cell as well.

KRas

The Ras family of genes is responsible for 20 percent of all cancers. They are oncogenes, that were originally proto-oncogenes. However, the most dangerous form is KRas. Kras, being a proto-oncogene, normally acts to help cell growth. However, overexpression of this gene eventually leads to the development of cancer. KRAS-4B is the primary isoform(variant)in human cancers, and it causes almost 90 percent of all pancreatic cancers, around 30 to 40 percent of colon cancers, and around 20 to 15 percent of lung cancers.

Most ways of treating this overexpression of KRas includes using inhibitors to block its activation, thus stopping it. However, there are many other ways that could be explored to tackle KRas.

Using Crispr to Combat KRas

Adenoviral Vectors

Did you just say a gutless virus?

Well, yes I did! Adenoviruses have gone through many generations of modification, but the most recent modification that we have made is to essentially wipe everything in their genome, except for the ITRs and a packaging signal.

Here the genome of adenovirus is shown. The normal genome is shown in black, and deletions are white. Insertions of transgenes are labeled in gray. https://www.nature.com/articles/3302612

By doing this, the gutless adenovirus also has been dubbed the helper-dependent adenovirus, as it needs another virus to replicate. To put it simply, another adenovirus will code for the whole entire virus(capsid, proteins, etc.) but it will not get it’s genome inserted inside as it has its packaging signal removed, thus allowing for it to not get packaged inside of a virus. If you want to learn more about these types of viruses, you can go here to check out my article on viral vectors as a whole.

Plasmid insert of DNA and gRNA that targets KRas. Generated via Benchling

Shown above, is how my gutless adenovirus will look like. It will contain the Cas9(Streptococcus Pyogenes)coding section, along with a section that will code for the KRas-targeting gRNA. This plasmid can be cloned either through polymerase chain reaction or through Gibson assembly.

Expected Results

Moreover, since this is a novel plasmid, it should be first tested in the lab in vitro(in glass/test tubes) before being used in clinical. trials.

Conclusion

Thank you for reading this! I am Sam, and I am a TKS innovator that is really passionate about seeing how we can leverage gene editing to help in future space travel! You can check out my other medium articles about tech and mindsets by clicking on my icon down below. You can contact me through my LinkedIn or my email: samlitks2@gmail.com. Thanks again, and have a wonderful rest of your day!

I’m a TKS activator and I’m fascinated in understanding the mind and exponential technologies and how understanding both can lead to discovery and innovation!

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