Synthetic Biology

Synthetic biology is an empowered genetic engineering discipline that comprises the design, fabrication and incorporation of synthetic DNA molecules into living organisms. Synthetic biology speeds up the design-build-test-learn cycle, solving long-standing biological research challenges.

A 16S RNA phylogenetic tree of Burkholderia. Species classically classified as pathogenic or beneficial are shaded in red or green, respectively. Blue and yellow diamonds denote outliers: species with also biotechnological interest or found to cause infections, respectively.

The genus Burkholderia is one group of bacteria for which their industrial use can benefit from synthetic biology.

The long-term goal of this research program is to unleash Burkholderia’s biotechnological potential by the development of synthetic biology tools. By identifying and characterizing Burkholderia genetic elements that control growth, survival, and energy production, we aim to engineer fully controllable Burkholderia strains.

Our laboratory has developed several genetic tools that can be applied in gene and genome editing to manipulate desired features in Burkholderia strains for growth control during biotechnological applications. The latest technology we developed is CRISPR-based interference (CRISPRi) for Burkholderia. CRISPRi for Burkholderia is available through Addgene.

Development of CRISPRi in Burkholderia. CRISPRi is a method of silencing native gene expression based on a catalytically inactive or dead endonuclease Cas9 (dCas9). In CRISPRi, a single guide RNA (gRNA) designed toward the 5′ end of the target gene and the dCas9 protein form a RNA-protein complex. The target region is recognized by the dCas9-RNA complex by base-pairing, sterically blocking transcription initiation. CRISPRi in Burkholderia.

In collaboration with Dr. David Levin, From Biosystems Engineering, U. of Manitoba, we are looking into the ability of Bcc bacteria to degrade polyesters used in the production of biodegradable plastics. Once the most active strains are identified, we will study the specificity of the bioplastic degrading enzymes and the genetic elements involved. The next step will be to edit the strain’s genome to produce a strain active regarding biodegradation, but defective in terms of pathogenicity.

Identification of genetic elements related to the extracellular degradation of polymers in Burkholderia strains. The high throughput of TnSeq enables parallel identification of thousands of mutants, empowering discovery of all possible contributing

We have developed a method for screening Burkholderia strains for bioplastic degradation on agar containing colloidal suspensions of plastics. By using large-scale transposon mutagenesis, we are identifying related genetic elements.

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