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CERES: CRISPR Engineering for the Rapid Enhancement of Strains

Technical Report ·
DOI:https://doi.org/10.2172/1820694· OSTI ID:1820694
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  1. Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
  2. Purdue Univ., West Lafayette, IN (United States)
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Previous strain development efforts for cyanobacteria have failed to achieve the necessary productivities needed to support economic biofuel production. We proposed to develop CRISPR Engineering for Rapid Enhancement of Strains (CERES). We developed genetic and computational tools to enable future high-throughput screening of CRISPR interference (CRISPRi) libraries in the cyanobacterium Synechococcus sp. PCC 7002, including: (1) Operon- SEQer: an ensemble of algorithms for predicting operon pairs using RNA-seq data, (2) experimental characterization and machine learning prediction of gRNA design rules for CRISPRi, and (3) a shuttle vector for gene expression. These tools lay the foundation for CRISPR library screening to develop cyanobacterial strains that are optimized for growth or metabolite production under a wide range of environmental conditions. The optimization of cyanobacterial strains will directly advance U.S. energy and climate security by enabling domestic biofuel production while simultaneously mitigating atmospheric greenhouse gases through photoautotrophic fixation of carbon dioxide.
Research Organization:
Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States); Sandia National Laboratories (SNL-CA), Livermore, CA (United States)
Sponsoring Organization:
USDOE National Nuclear Security Administration (NNSA)
DOE Contract Number:
NA0003525
OSTI ID:
1820694
Report Number(s):
SAND2021-11486; 699441
Country of Publication:
United States
Language:
English

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09 BIOMASS FUELS
59 BASIC BIOLOGICAL SCIENCES
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