Development and use of novel inducible Cas9 models to study gene function

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Krishnamoorthy, Vignesh
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CRISPR/Cas9 based mouse models are gaining traction for in vivo disease modeling. However the existing models do not allow both the spatial and temporal control of Cas9 expression which may lead to off-target effects. This thesis employed two different approaches to utilize/develop Cas9 models to replicate tumorigenesis with reduced off target effects. The first approach illustrated the utility of a drug inducible Cas9 model to recapitulate sporadic tumor development when Cas9 is induced along with the delivery of gRNAs targeting desired genes. In the second approach, we have attempted to develop a mouse model wherein both gene inactivation and gene activation could be achieved by creating an ES cell model with both Cas9 and a gene activation complex. ES cells expressing conditional and inducible variants of Cas9 and the gene activation complex has been created and initial data suggests that the system should be expressing Cas9 and the gene activation complex.
Mouse modeling, Tumor modeling