CRISPR/Cas9 to repair RDEB gene

 

 

 

 

 

 

 

• Project summary (top of page)
• About our funding
• Latest progress summary
• About our researchers
• Why this research is important
• Researcher's abstract
• Researcher's progress update

  

About our funding:

Research leader	Dr Sergio López-Manzaneda
Institution	Fundación Jiménez Díaz, CIEMAT, Spain
Type of EB	DEB
Patient involvement	No
Funding amount	£15,000
Project length	1 year
Start date	TBC 2024
DEBRA internal ID	GR000042

 

Latest progress summary:

Due 2025.

 

About our researchers:

Lead researcher:

Dr Sergio López-Manzaneda is a researcher at CIEMAT with a strong background in genome editing.

Co-researchers:

Dr Fernando Larcher is associate professor of biochemistry at Universidad Carlos III de Madrid and Head of Division at CIEMAT (Epithelial Biomedicine Division).

Alex Bassons Bascuñana is a PhD student in the Epithelial Biomedicine Group of the Biomedical Innovation Unit at CIEMAT.
 

Collaboration: 

Dr Paula Rio is an internationally recognised expert in gene editing and gene therapy of inherited hematopoietic diseases, from basic science to clinical trials.

 

Why this research is important:

This non-viral gene editing approach looks for a low-cost and universal strategy to generate a library of tools capable to address every known mutation... In future pre-clinical studies, gene “patched” edited keratinocytes and fibroblasts from patients will be used to generate bioengineered skin equivalents.

Dr Sergio López-Manzaneda

 

Researcher’s abstract:

Grant title: Non-viral genome editing patching of COL7A1.

In this project, we propose a genome editing strategy focused on the treatment not of just one point mutation but groups of neighboring mutations causing RDEB, by repairing full exons harboring them. This “genetic-patching” is based on the natural Homologous Recombination (HR) DNA repair mechanism facilitated by the CRISPR/Cas9 system. The “patches” represent small (300-400 base pairs) double stranded HDR donor DNA templates that would be designed from gene sequences enough to cover few contiguous exons, in this case, corresponding to the COL7A1 gene encoding type VII collagen. Our aim is to characterize (efficacy, safety) and to standardize the use of these technology with the idea of having specific patches readily available to tackle individual groups of mutations. The non-viral technology proposed for ex vivo RDEB patient cell treatment will facilitate its translation to the clinic at low cost. 

One of the most promising therapeutic approaches for EB is genome editing using CRISPR/Cas9 technologies. During the last 7 years our laboratory has been actively working in this field providing strong proof-of-concept data aiming at translating its results to the clinic. While we are in the verge of clinical application with one of these approaches that obtained an Orphan Drug designation by the European Medicines Agency (EU/3/20/2253), we continue seeking for more efficient, safer and clinically relevant genome editing strategies.

In this project, we propose to test an innovative gene editing approach that does not require the use of viral vectors (all material is introduced by a single electroporation). The goal is to use these “genetic patches” to correct mutated regions after having cut this genetic region with CRISPR/Cas9 technology. We have designed four different “patches”, this means, dsDNA donor templates specifically modified in their ends by a proprietary technology of our provider (Integrated DNA technologies, IDT) to favour HDR. These templates contain two exons and their surroundings (73 and 80, two “patches” each) and four different cut points (two for Exon 73 and two for exon 80). This templates  (around 300-400 bp) also cover the neighbour exons.

We aim to evaluate the extent of the repair within the “patches” which would allow us to address groups of mutations or, if the system works well enough, to fully correct the exons within the “patches”. We believe that this novel, no viral, personalized gene editing strategy could be easily translated to the clinic with a significant low economical cost.

 

Researcher’s progress update:

Due 2025.

 

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