Til bevillingsoversigt

Selective and traceless release of truncated CRISPR Cas9 guide RNA for anti-cancer gene editing in NSCLC cells

Carlsbergfondets internationaliseringsstipendier

What

In this project, the gene editing CRISPR/Cas9 system will be used to disable lung cancer cells in a laboratory setting. The gene editing will be directed by a synthetic guide RNA sequence that will be optimised by comparing the effect of many such compounds with subtle modifications. When several highly active RNAs have been developed, they will be conjugated to a carrier system that will ensure delivery to cancer cell. The carrier system is based on cell penetrating peptides that will be optimised to target the cancer cells. The final conjugates are expected to deliver more than 50% of the RNA in a 24 period and suppress the cancer cells at least three times more effective than current guide RNA.

Why

Gene editing holds huge promise as treatment for human diseases such as cancer. In order to realize this potential, several challenges first have to be solved. Ensuring that the developed drugs reach the cancer cells and the correct targets within the cells is one of those challenges. This is challenging because the bimolecular drugs are relatively large and unstable compared to traditional small molecule drugs that require less or no assistance in reaching their site of action. This project can provide new tools to direct bimolecular drugs for gene edition and is therefore an important part of the further development of gene editing technology as a whole.

How

The experimental plan for this project will begin by optimizing the guide RNA that directs the gene editing. The RNA will be chemically modified to ensure stability and increase function. Sequencing will be used to show that the targeted cancer genes are successfully disabled and to what extent off-target genes are damaged. Next, the carrier peptides will be optimized to transport the guide RNA into lung cancer cells. Fluorophores will be used to track the compounds and visualize how they enter the cell. Once suitable pairs of guide RNAs and peptides have been identified, they will be connected in such a way that the RNA can be released inside the cell and exert the cancer disruptive function.

SSR

The impact that gene editing will have on society is hard to predict but is likely to be significant. Because of this, it is important to establish safe and reliable methods that will allow gene editing to be used responsibly. Cancer likewise has a large impact on society and still require new developments in treatment. This project has the potential to create new knowledge in areas and can therefore eventually lead to direct value for society.