Cellecta, Incorporated — Department of Health and Human Services SBIR Phase II: 172

Description

Open Resource CRISPR CASGenome Wide sgRNA Library Screening PlatformABSTRACTCRISPR technology made its giant leap from bacteria to mammalian system when a few landmark discoveries that demonstrated Casprotein from Streptococcus Pyogenes could be reprogrammed with synthetic RNAssgRNAsto generate site specific double strand breaksDSBsin vitro and in mammalian cellsCRISPR technology appealed to the world of genomics and gene editing as it made the task of generating large genome editing libraries possible for functional genetic screensAlthough there has been a tremendous increase in CRISPR related publications over the last two years in many areas of drug discovery and gene function analysisit is already being recognized by the scientific community that the application of the technology to high throughput functional genomics screening comes with its own unique challenges and a significant improvement is warrantedAs research in this field focuses on improving effectivenessincreasing selectivity and reducing offtarget effectscompanies aim to develop simplifiedflexiblerobust and cost effective CRISPR screening platformsincorporating all of the above with experimental and bioinformatics tools for data validation and integration of this information into operational cell based modelsCRISPR technology can be used to either achieve sgRNA directed gene knockout by CasendonucleaseCRISPR KOor sgRNA directed regulation of gene expression by mutant dCasCRISPRa for transactivationCRISPRi for inhibitionWhile published studies have provided proof that both CRISPR KO and CRISPRa i can be successfully used in high throughput functional genomics screenssubstantial margins of improvements do exist in multiple aspects of the technologyAdditionallyalthough the CRISPR system has opened many potential avenues for improving the drug discovery processthose remain only potential opportunities until we develop robust commercially available CRISPR screening technologiesas well as experimental and bioinformatics tools for data validation and integration of this information into operational cell based modelsFollowing the successful completion of Phase I studiesin the present Phase II application we propose the final development and commercialization of a high performance CRISPR screening platform with innovative features enhancing the frequency of effective gene knockout in CRISPR KO applicationsand the intensity of promoter activation repression in CRSPRa iAs supporting toolswe will provide protocolsreagents and software tools for screening data analysis and validationThe ultimate goal of this project is to developvalidate and commercialize a set of high efficiencyreadyto screengenome wide pooled sgRNA human and mouse CRISPR lentiviral librariessupporting bioinformatics tools for data analysisas well as custom services for CRISPR library designCRISPR functional genetic screeningand hit confirmation validation assaysUpon completion of the proposed Phase II studiesCellecta will be ready to provide the scientific community with a comprehensivehighly efficientscalable and costeffective genetic screening tool set that is expected to expedite systematic identification of new targets for therapeutic intervention and facilitate the development of highly specific drugsbiomarkers and novel therapeutic conceptsThe new CRISPR screening platformincluding sgRNA librariescustom servicessupporting software and data set tools developed in the course of Phase I II studies will be made available to all academic and commercial researchers through www cellecta com web site