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High Throughput Tool Development for Non-Conventional Organisms
High Throughput Tool Development for Non-Conventional Organisms
The most well-studied species have provided important insight into molecular genetics. However, engineering of these organisms has limitations that could be overcome by rapidly engineering tools for new organisms. One emphasis of our lab is to fill this gap by developing genetic engineering tools for species that can provide new insights, produce valuable metabolites, or that can survive in extreme conditions like high salt or high temperature. For example, we are engineering Marchantia polymorpha to rapidly understand plant genetics and screening hundreds of strains of Kluyveromyces marxianus to find extremophile yeast for production of valuable chemicals.
CRISPR-Cas9 Genome Wide Screens
CRISPR-Cas9 Genome Wide Screens
Forward genetics helps uncover the role of genes and can be used to engineer organisms to produce valuable chemicals. CRISPR-Cas9 uses a gRNA barcode to cut a single gene in an organism. One tool that our lab develops are CRISPR Cas9 genome-wide screens in non-conventional hosts. By creating gRNA barcodes for every gene in the genome of a species, we create a pool of single knockout mutants. This helps us understand less understood species and how their genetics can be used to help us create biochemicals in a more sustainable way.
Key Publication: Robertson et al., Metabolic Engineering, 2024
Robertson et al., bioRxiv, 2024
Beltran et al., Nature Biotech, 2022
Biosensor Development Using Repurposed Plant Hormone Receptors
Biosensor Development Using Repurposed Plant Hormone Receptors
Designing sensitive, specific, and portable biosensors remains a challenge in biotechnology. We have repurposed PYR1 (Pyrabactin Resistance 1), a plant abscisic acid (ABA) receptor, as a chemically induced dimerization yeast 2-hybrid biosensor platform. This system has proved to be amenable to a wide variety of ligands, portable to multiple hosts, and useful for many different applications. Including rapid biosensor screening in S. cerevisiae, biosensor coupled CRISPR Cas-9 genome wide screens in K. marxianus, and environmental sensing in A. thaliana. We continue to advance this system to expand its ligand targets and develop new uses.
Key Publication: Beltran et al., Nature Biotech, 2022
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