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US 10,329,587 B2Granted
Orthogonal Cas9 proteins for RNA-guided gene regulation and editing
AssigneePresident and Fellows of Harvard College
InventorsGeorge M. Church, Kevin M. Esvelt, Prashant G. Mali
Life of this patent
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Abstract
Methods of modulating expression of a target nucleic acid in a cell are provided including use of multiple orthogonal Cas9 proteins to simultaneously and independently regulate corresponding genes or simultaneously and independently edit corresponding genes.
Claims17
A cell in culture comprising
- a first foreign nucleic acid encoding two or more guide RNAs complementary to two or more respective target nucleic acids,
- a second foreign nucleic acid encoding two or more orthogonal RNA guided nuclease null DNA binding proteins, wherein each of the two or more orthogonal RNA guided nuclease null DNA binding proteins is of a different species and recognizes a corresponding PAM sequence in a target nucleic acid, and
- a third foreign nucleic acid encoding two or more transcriptional regulator proteins, wherein the cell is configured to express the two or more guide RNAs, the two or more orthogonal RNA guided nuclease-null DNA binding proteins, and the two or more transcriptional regulator proteins, wherein the cell comprises two or more co-localization complexes with each including a guide RNA, an orthogonal RNA guided nuclease null DNA binding protein, a transcriptional regulator protein and a target nucleic acid.
Dependencies are inferred from the claim text.