Extracellular vesicles encapsulating CRISPR machinery for treatment of SARS-CoV-2 infection

NIH Pandemic-Era Grants

Pandemic Era Grants

2024

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Principal Investigator: Houjian  Cai
Organization: UNIVERSITY OF GEORGIA
Fiscal Year: 2024
Award: $184,222
Funding agency: National Institute of Allergy and Infectious Diseases

Summary/Abstract
SARS-CoV-2 has caused the deaths of millions of people globally. Effective antiviral therapeutic
treatment options are urgently needed. CRISPR-mediated genome editing has provided a very promising avenue
for treatment of a variety of genetic diseases. Particularly, the CRISPR-Cas13 system has been demonstrated
to possess the potential of inhibiting SARS-CoV-2 and influenza infections by degradation of viral genomic RNA
and viral mRNA. However, it is still very challenging to deliver the CRISPR machinery to initiate genome editing
efficiently in vivo. Extracellular vesicles (EVs) contain a variety of molecular components including lipids, mRNA,
microRNAs, and proteins. A large body of studies has shown that EVs mediate cell-to-cell communication by
transmitting their encapsulated contents. This proposal intends to construct EVs encapsulating the CRISPR
machinery and deliver the EVs to respiratory epithelial cells to inhibit SARS-CoV-2 proliferation in vivo. We will
therefore genetically engineer the Cas13d protein so that Cas13d/CRISPR-RNA (crRNA) ribonucleoprotein
complex can be encapsulated into EVs. We will also engineer the membrane of EVs, such that EVs target
respiratory epithelial cells and deliver Cas13d/crRNA for inhibiting SARS-CoV-2 viral assembly and proliferation,
thereby inhibiting COVID-19. This study will allow us to understand the feasibility of an EVs-based vehicle to
deliver genome editing machinery to inhibit SARS-CoV-2 proliferation in lung epithelial cells. This study will
provide a treatment option for COVID-19 patients to reduce disease severity and mortality.

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