Developing experimental treatments to eliminate the defective gene in KRT6A using the CRISPR/Cas technique
In a previous study in our laboratory, we developed a model of the disease using cells from patients with a specific genetic mutation. This allowed us to recreate the effects of the disease in a humanized mouse skin model. Gene therapy is the most promising strategy for treating this disease, as it aims to deactivate the mutated allele of the gene, which negatively affects the healthy allele. Traditional therapies, which attempt to introduce a functional copy of the gene, are not effective in this case.
Thanks to advancements in genetic editing technology and using a molecular tool known as CRISPR/Cas, we can now explore ways to specifically remove the mutated allele of the gene. These “molecular scissors” only activate and remove the target allele under specific conditions.
We discovered that a particular mutation in the KRT6A gene, which is affected in this disease, creates the right conditions to activate the scissors and remove only the mutated allele.
We have tested this technique on patient cells in the laboratory with successful results: we were able to correct the gene and restore the skin’s structure. Currently, our goal is to find a way to directly introduce these tools into the patients’ cells, without the need to extract, correct them in the lab, and then transplant them back to the patient. To achieve this, we are working on developing the best delivery vehicle that can carry these tools directly into the patients’ skin, thus solving the problem in the mutated cells.
Note: What is an allele?
Each gene contains the instructions necessary to perform a specific function in the organism. Each person inherits two alleles for each gene, one from the mother and one from the father. For example, the gene responsible for eye color can have alleles that code for brown, blue, or green eyes. These alleles can be identical (for example, both alleles code for brown eyes) or different (for example, one allele codes for brown eyes and the other allele codes for blue eyes). The combination of alleles a person has for a particular gene influences their visible or functional traits.
In the case of certain diseases, such as in the skin diseases, the K6a gene, for example, may have a healthy allele that provides the correct instructions for making the K6a protein, and a mutated allele that codes for a defective protein. In this context, the mutated allele tends to be dominant over the healthy allele, meaning that even though the person produces some functional protein from the healthy allele, the defective protein produced by the mutated allele affects the overall function. This leads to the manifestation of the disease. The goal in these cases is to eliminate the mutated allele so that the healthy allele can effectively produce the correct protein, thus correcting the dysfunction and potentially curing the disease.
To learn more about Marta’s project, funded by PC Project, click here.
