The problem our project solves
Coronavirus 2 of severe acute respiratory syndrome (SARS-CoV-2) belongs to the Coronaviridae family of viruses. This family of viruses is characterized by having its genome formed by a long positive single-stranded chain of RNA. The CRISPR system (Clustered Regularly Interspaced Short Palindromic Repeats) is proving to be an exceptional tool for genome editing and whose potential is constantly increasing. The CRISPR system and its associated genes (Cas) is present in both archaea and bacteria and constitutes an adaptive immune system directed by RNA, this system is useful in defense against viruses and other pathogens. One of the first applications of Cas13 that is being explored is its use to be directed against RNA virus genomes, even more recently against SARS-CoV-2 itself, and that could be implemented as a therapeutic tool against these viruses. Despite the versatility and simplicity of the CRISPR system, its use in vivo is limited to its effective administration to reach target cells. Therefore, we propose to nanoencapsulates Cas13d systems to deliver gene-editing tools targeting SARS-CoV-2, as a platform for the administration of therapeutic CRISPR / Cas13d tools in cells infected of COVID-19 patient.
The solution we bring to the table
For the delivery of CRISPR systems is required the transport of gene editing components directly into the nucleous of targeted cells. The delivery system that is being proposed SpherAQ ™, can combine the advantaged given by the 100% multilayer biodegradable and biocompatible shell and the specific structure of the particle characterized with their small size. These features guarantee transparency, higher solubility in solution, without the presence of agglomeration and coalescence phenomenon, and last but not least- higher bioavailability. The technology used to produce the capsules is mild and does not involve the use of thermal treatment that might affect the integrity of Cas13d systems. Cellular uptake of SpherAQ ™ was demonstrated in several types of cells like Caco2, HeLa, HepG2 and J774A1 macrophages.
What you have we during the weekend
During this amazing weekend, we were able to create a multidisciplinary team with people having a different background, all very valuable and able to contribute to this project. We have learnt a lot from each other, both when it comes to nanoencapsulation and Cas13d systems used against SARS-CoV-2. We have created an innovative therapy and a plan how to continue.
The solution’s impact to the crisis
Although the respiratory disease caused by SARS-CoV-2 is the main problem for COVID-19 patients and its airway presence is used to confirm the clinical diagnosis, the virus is detected at a similar level to nasopharyngeal swabs, in other sites, such as feces. Additionally, the receptor used by SARS-CoV-2 as entrance in the patient cells, ACE2, is widely expressed in different cell types, such as lung alveolar cell type 2 or colon cells. This suggests that other organs might be reservoir for the virus. Consequently, the potential SARS-CoV-2 transmission might occur through other ways than respiratory ways. In order to reduce virus burden in the patient’s intestine to favor the complete elimination of the disease and to reduce the risk of dissemination of the virus, we propose used encapsulated Cas13d system to target SARS-CoV-2 in the intestine tract. SpherAQ ™ can be produced in in liquid form with the possibility to be put in aerosol for lung administration, or as powder or granules for oral administration as the formulation reaches the intestine. The nanoencapsulates can be produced in a range of 25-800 nm, charged, and the particle itself can be layered giving it different functionalities and tailoring it to perfection, making the formulation versatile and easily adaptable for any necessity. Since it is encapsulated it has a superior stability and can be transported and stored in an easy way.
The necessities in order to continue the project
Our project links two innovative technologies, such as gene editing and nanoparticle technology, in order to complement each other. The members of our team have the expertise in both technologies and the symbiosis among them will facilitate the right development of the project. Despite the experience of the members of team, the project will require additional personal and budget, since none of the groups have been previously focused on coronavirus field. Additionally, we have realized that to achieve the project, we need to add a group specialized in Coronavirus biology to determine more precisely the region of genome of SARS-CoV-2 to target by the encapsulated CRISPR/Cas13d system. Additionally, we should include any research or clinical group with Biosafety Level 3 Laboratory to be able to manipulate infected cells and animal models, such as the commercially available K18-hACE2 mice to evaluate the efficacy of our solution in vitro and in vivo. If these needs are met, our project could be transferred to a clinical trial to assess the efficacy of the encapsulated Cas13d system in COVID-19 patients.
The value of our solution after the crisis
CRISPR system is easy to design and can target any kind of sequence of interest, therefore the applicability of this strategy is limitless. Seen as one of the major issues related to virus infection is their ability to quickly mutate in order to avoid the host’s defense system, the discovery of a cure or vaccine would result temporary. CRISPR technology could be the perfect weapon, enabling us to respond in a fast and adaptive way to changes of the virus.