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US 11,306,308 B2Granted
High-throughput CRISPR-based library screening
AssigneesMassachusetts Institute of Technology, The Brigham and Women's Hospital, Inc.
InventorsDavid K. Gifford, Nisha Rajagopal, Richard Irving Sherwood
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Abstract
Provided herein is an improved method for performing CRISPR/Cas based screening that is not dependent on viral cloning methods.
Claims14
A multiplexed editing regulatory assay (MERA) construct comprising:
- one or more copies of a defective gRNA expression construct wherein each defective gRNA expression construct comprises:
- i. a dummy gRNA that is not homologous to a eukaryotic genome, and is unable to target a Cas nuclease to the genome for a double strand break,
- ii. a guide hairpin sequence,
- iii. a native regulatory region which will be targeted using CRISPR/Cas endonuclease;
- and
- iv. an operably linked promoter.
3 more elements
Dependencies are inferred from the claim text.