Expanding the functional metagenomics toolkit with CRAGE and CRISPR-Cas9 in taxonomically diverse microbial host chassis
EMSL Project ID
50967
Abstract
Plant and microbial biomass offers a renewable alternative to energy and materials produced from oil. The industrial scale production or biorefining of sugars and aromatic compounds from biomass needs more effective and efficient biocatalysts. For almost 4 billion years microbial communities have been driving energy and material transformations that create and sustain life on our planet. As a result, although the vast majority of microbes in nature remain uncultivated, they represent a deep reservoir of genetic information and metabolic potential. Functional metagenomics screening can access the hidden metabolic powers of microbial communities to recover biological devices from environmental genomes for sustainably deconstructing plant biomass into energy and materials. Here we propose to further develop genetic toolkits for enzyme discovery, metabolic engineering and gene function characterization, for biocatalysis and the biosynthesis or bioproducts from renewable feedstocks. We will leverage the capacity of JGI and EMSL, and the chassis-independent recombinase-assisted genome engineering (CRAGE) system established at the JGI, to develop enabling CRISPR-Cas9-based tools and functional screening chassis, including E. coli EPI300, Arthrospira platensis (Spirulina) and Synechococcus elongatus, that can be used to both discover new biocatalysts and to funnel biomass into renewable value-added products. This approach requires a hybrid analytic capacity that can be directly provided in collaboration with the JGI and EMSL user facilities with respect to integrated sequencing, proteomic and metabolomic analyses.
Project Details
Project type
FICUS Research
Start Date
2019-10-01
End Date
2022-08-31
Status
Closed
Released Data Link
Team
Principal Investigator