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US 10,815,467 B2Granted
Simultaneous multiplex genome editing in yeast
AssigneeInscripta, Inc.
InventorsMiles Gander, Tian Tian, Skylar Stefani
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Abstract
The present disclosure provides compositions of matter, methods and instruments for editing nucleic acids in live yeast cells.
Claims20
A composition of matter for performing simultaneous multiplex RNA-directed nuclease editing in yeast comprising from 5′ to 3′:
- a pol II promoter;
- a transcription start site;
- a first editing cassette wherein the first editing cassette comprises a coding sequence for a first gRNA covalently linked to a coding sequence for a first donor DNA, wherein the first donor DNA comprises a rational, desired edit to a first target sequence and an edit which is able to render inactive a first proto-spacer motif (PAM) in the first target sequence;
- a linker;
- a second editing cassette wherein the second editing cassette comprises a coding sequence for a second gRNA covalently linked to a coding sequence for a second donor DNA, wherein the second donor DNA comprises a rational, desired edit to a second target sequence and an edit which is able to render inactive a second proto-spacer motif (PAM) in the second target sequence;
- a coding sequence for a self-cleaving ribozyme;
- and a pol II terminator.
4 more elements
Dependencies are inferred from the claim text.