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US 11,401,530 B2Granted
Bacteriophage-based artificial viruses for human genome remodeling
AssigneeThe Catholic University of America
InventorsVenigalla B. Rao, Jingen Zhu
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Abstract
Described is an “artificial virus” (AV) programmed with biomolecules that can enter human cells and carry out precise human genome modification. The AVs comprise: at least one viral vector, such as bacteriophage T4; at least one therapeutic molecule, such as DNA, RNA, protein and their complex; and a lipid coating. Also described is a method of human genome modification, using such an AV, and a method of program such an AV.
Claims3
A CRISPR-based method of programming artificial virus (AV) with genome modification capabilities comprising:
- generating a “acceptor” phage by deleting ipI and ipII genes from a wild type T4 phage;
- generating a host bacteria cell with a plasmid containing a gene of target protein and a spacer plasmid that expresses Cas9 or Cpf1 and CRISPR RNA corresponding to a protospacer sequence in the deleted region of the acceptor phage;
- infecting the host bacteria cell with the “acceptor” phage;
- recovering an engineered “acceptor” phage from the host bacteria cell;
- obtaining an empty engineered T4 capsid from the engineered “acceptor” phage;
- packaging at least one DNA in the engineered T4 capsid,
- wherein the gene of target protein is flanked by capsid targeting sequence (CTS) at the C-terminus and nuclear localization sequence (NLS) at the N-terminus to form CTS-gene-NLS
- sequence.
5 more elements
Dependencies are inferred from the claim text.