Mesophilic argonaute systems and uses thereof
AssigneesThe Board of Trustees of the Leland Stanford Junior University, Intima Bioscience, Inc.
InventorsLei S. Qi, Modassir Choudhry, Xueqiu Lin, Trevor N. Collingwood, Thomas Henley, Benjamin Klapholz, Tilmann Buerckstuemmer, Sejla Salic
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Abstract
Constructs comprising Argonautes and neighboring genes are disclosed for use in gene editing. Disclosed are also compositions and methods utilizing these Argonautes and neighboring genes. Also disclosed are the methods of making and using the Argonautes and neighboring genes in treating various diseases, conditions, and cancer.
Claims33
1-63. (canceled)
A recombinant fusion polypeptide, said fusion polypeptide comprising:
- (a) an Argonaute (Ago) polypeptide, wherein said Ago polypeptide is a Clostridia Ago (C-Ago) polypeptide;
- and (b) a nucleic acid unwinding polypeptide, wherein upon contacting a target polynucleic acid with the fusion polypeptide in the presence of a guiding polynucleic acid, the fusion polypeptide cleaves the target polynucleic acid.
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A system comprising:
- a. a mesophilic Clostridia Argonaute (C-Ago) polypeptide, or a polynucleic acid encoding the same, or a functional fragment or variant thereof;
- and
- b. an exogenous non-naturally occurring guiding polynucleic acid that binds a target polynucleic acid,
- wherein the target polynucleic acid is a single stranded DNA (ssDNA), a double stranded DNA (dsDNA), a single stranded RNA (ssRNA), or a double stranded RNA (dsRNA), and wherein upon contacting the target polynucleic acid with the system at a mesophilic temperature, the system cleaves the target polynucleic acid.
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A recombinant fusion polypeptide, said fusion polypeptide comprising:
- a. a mesophilic Argonaute (Ago) polypeptide;
- and
- b. a single strand DNA binding protein (SSB).
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149-160. (canceled)
Dependencies are inferred from the claim text.