Using CRISPR as an activator to target promoters of long non-coding RNAs

Application

• A gene therapy approach where complex loci can be activated through the use of CRISPR resulting in the upregulation of noncoding RNAs.
• The use of vectors, such as adenoviral vectors, for the delivery of CRISPR components allows the application of this strategy to multiple cellular contexts and various therapeutic applications

Development Status

• Proof-of-concept, in vitro data.

IP Status

• Patent application no. WO2023242545A1 has been published 21 December 2023.

Commercial Offering

• Co-development or licensing partner.

Opportunity

The technology has been utilised in the context of pathological vascular remodelling following injury to the vasculature, a major unmet clinical need. Vascular smooth muscle cells (vSMCs) have a crucial role in pathological vascular remodelling, and targeting vSMCs may hold significant promise for treating cardiovascular diseases.

A key factor regulating cellular states, e.g., vSMC identity, is the synergistic expression levels of noncoding RNAs, long noncoding RNAs (lncRNAs) and microRNAs (miRNAs). Complex noncoding genomic loci can have multiple promoters and include different classes of noncoding RNA molecules.

There are several strategies to enhance RNA expression of genomic loci, however (1) the transcript size might exceed the vector limit; (2) the locus could include multiple transcripts which cannot be simultaneously co-expressed; (3) the exact structure of the locus might not be well annotated.

Technology Overview

Elevating the expression of a large lncRNA axis, as opposed to overexpressing its individual components, presents a preventive effect on the development of vein graft failure. This is done by utilising CRISPR activator technology to activate the promoter region of noncoding RNA axes, CARMN and MIR503HG. Both lncRNAs have been shown to be downregulated in different pathological contexts involving primary vascular cells, therefore the activation of their expression is important to maintain cell identity.

Our results show that it is possible to activate the expression of the lncRNA transcripts concomitantly to microRNAs by activating the main promoter of the noncoding RNA axes. We also have shown that it is possible to use this strategy in primary smooth muscle cells opening the possibility for translation to a clinical setting.

Benefits

• Treatment for diseases that have a substantial unmet clinical need. Current treatment strategies are more challenging and highly risky.
• Single dose cure - requires only one intervention and one vector.
• A low-cost strategy for global activation of the whole set of transcripts within a locus.
• The use of adenoviral vectors for CRISPR delivery allows the application of this strategy to multiple cellular contexts and allows therapeutic transability.

Quote: TEC1104437