|United States Of America||Published Application||20190194632||06/27/2019||2016-132|
|Patent Cooperation Treaty||Published Application||WO2017189336||11/02/2017||2016-132|
Many advancements to the Cas9 system
(both the Cas9 nuclease and the sgRNA sequence) have been made to increase and
optimize its efficiency and specificity. Since many diseases and traits in humans have
a complex genetic basis, multiple genomic targets must be simultaneously edited in order for
diseases to be cured or for traits to be impacted. Thus in order for CRISPR/Cas9 to be an
effective gene therapeutic technology, huge swathes of the genome must be
edited simultaneously, efficiently, and accurately.
To address many of these issues, UC Berkeley researchers have developed a system method to rapidly manipulate multiple loci. This system allows for either sequential (maintaining inducible Cas9 present in the genome) or simultaneous (scarless excision) manipulation of Cas9 itself and can be applied to any organism currently utilizing the CRISPR technology. The system can also be applied conveniently to create genomic libraries, artificial genome sequences, and highly programmable strains or cell lines that can be rapidly (and repeatedly) manipulated at multiple loci with extremely high efficiency.