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Improvements to Cas9-Mediated Mutation

Technology Benefits
High efficiencyTargeted mutagenesisNo off-targets with dCas9 editing
Technology Application
Precision gene editing in therapeutic contextsIntroduction of desired traits in cropsGeneration of genetically modified organismsIsogenic cell line generation
Detailed Technology Description
None
Others

Publication


Enhancing homology-directed genome editing by catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA


Additional Technologies by these Inventors


Tech ID/UC Case

25181/2016-001-0


Related Cases

2016-001-0

*Abstract

Cas9 is an RNA-guided DNA endonuclease used to perform targeted genomic manipulations, which can include the error-prone knockout of sequences via non-homologous end joining (NHEJ) and the introduction of precise edits via homology directed repair (HDR). HDR editing shows great promise for a variety of uses, such as generating new cellular immunotherapies, curing genetic disease, and introducing traits into agricultural crops. Yet the efficiency of HDR has lagged behind that of NHEJ, complicating these exciting applications. Additionally, worries have arisen about unintended knockout from off-target NHEJ.

 

UC Berkeley researchers have found that Cas9 operates by a surprising mechanism, which suggested ways to improve HDR. Taking advantage of this mechanism, researchers found simple methods to dramatically increase the efficiency of HDR, introducing targeted mutations in human cells with frequencies around 60%. Additionally, catalytically inactive Cas9 can be used to make mutations via HDR without attendant error-prone NHEJ. This latter activity allows the precise introduction of mutations with no danger of undesired knockout at off-target sequences.

 

*Principal Investigator

Name: Jacob Ellery Corn

Department:


Name: Christopher Richardson

Department:

Country/Region
USA

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