Prime editing to correct disease causative mutations in patient-derived Gaucher iPSC lines
(2023) MOBN03 20222Degree Projects in Molecular Biology
- Popular Abstract
- Prime editing to correct disease causative mutations in patient derived Gaucher iPSC lines
Lysosomes are the recycling bins of our cells, but what happens if they stop working? Do we get sick? Why would they stop working? Dysfunctions in lysosomes can be due to genetic mutations, changes in our genome that can lead to brain disease and premature death. Recently, the CRISPR/Cas9 system, that works like a pair of molecular scissors to modify the genome, was created and rewarded with a Nobel Prize. This genome editing tool will help us fight back against these life-threatening disorders and investigate how they cause disease.
Lysosomal storage disorders (LSDs) are a group of metabolic diseases caused by dysfunction of the recycling... (More) - Prime editing to correct disease causative mutations in patient derived Gaucher iPSC lines
Lysosomes are the recycling bins of our cells, but what happens if they stop working? Do we get sick? Why would they stop working? Dysfunctions in lysosomes can be due to genetic mutations, changes in our genome that can lead to brain disease and premature death. Recently, the CRISPR/Cas9 system, that works like a pair of molecular scissors to modify the genome, was created and rewarded with a Nobel Prize. This genome editing tool will help us fight back against these life-threatening disorders and investigate how they cause disease.
Lysosomal storage disorders (LSDs) are a group of metabolic diseases caused by dysfunction of the recycling centre of the cell, the lysosome. Many LSDs affect the brain of young children, and can lead to premature death. Gaucher Disease (GD) is the most common LSD, and the focus of this project. To date, there is no treatment available to treat neurological symptoms, and there is a lack of available human samples to study brain-related disease. However, we can make use of advances in human cell modelling to examine the impact of the mutations by using patient-derived induced pluripotent stem cells (iPSCs), stem cells that have been reprogrammed from fibroblasts without the need of human embryos.
Our lab has already established disease and healthy control iPSC lines, but due to differences in the genome of different people, we require healthy “copies” of the patient cells without the mutation to be certain of the impact of the disease-causative mutation. Therefore, our aim was to correct the mutations in patient-derived iPSCs using the aforementioned CRISPR/Cas9 system.
Due to difficulties in correcting these mutations with the normal CRISPR/Cas9 system, which works by breaking the DNA molecules at the region of interest, we optimized Prime Editing, a new genome editing tool derived from CRISPR/Cas9 that does not break the DNA. The advantage of this system is its safety to introduce the specific change of interest without generating unwanted DNA changes. With this approach, we were able to correct the disease mutations in two GD patient iPSC lines. These lines will be essential in the study of GD and are already being useful to understand the impact of a precise mutation. As stem cells can be differentiated into various cell types, it is interesting to use the 2 isogenic lines that I have produced by differentiating them into induced neurons or astrocytes, two important brain cell types or into brain organoids, mini brains produced in the lab, to understand what problems are occurring within the brain cells of patients.
It is difficult to treat neuronopathic disease due to blood brain barrier, a thin protective layer that block macromolecules, like drugs, to enter the brain. Gene therapies are ongoing to treat GD, however, they have large risk of causing side effects. Gene editing could therefore be a safer and more precise option to cure GD in the future. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9140665
- author
- Girard, Aline
- supervisor
- organization
- course
- MOBN03 20222
- year
- 2023
- type
- H2 - Master's Degree (Two Years)
- subject
- language
- English
- id
- 9140665
- date added to LUP
- 2023-10-31 12:06:47
- date last changed
- 2023-10-31 12:06:47
@misc{9140665,
author = {{Girard, Aline}},
language = {{eng}},
note = {{Student Paper}},
title = {{Prime editing to correct disease causative mutations in patient-derived Gaucher iPSC lines}},
year = {{2023}},
}