Dyadic scientists have developed and patented strains of the fungal microorganism Chrysosporium lucknowense – or C1 – to rapidly discover and express both eukaryotic and prokaryotic genes; and to manufacture the novel products of those genes, using C1 as the host organism from beginning to end. By utilizing a single organism for all steps of the process, Dyadic’s integrated platform eliminates many of the bottlenecks inherent with other technologies currently used to bring gene products to market.

Dyadic’s novel technology platform consists of two parts:
- The C1 Fungal High-Throughput Robotic Screening System (More at HTRS)
- The "C1 Express" Hyperproducing Protein Expression System (More at Gene Expression)

Dyadic’s One Stop Shop
By integrating these two technologies, Dyadic has created a "one-stop shop" for gene discovery, gene expression, product development, and product manufacturing. The process is outlined below:

Gene expression libraries are generated
DNA from various sources including individual organisms, environmental samples, or collections of genes and gene variants can be fragmented and cloned into Dyadic’s specialized C1 expression vectors. The resulting collection of cloned genes – which can number in the thousands – are transferred into C1.

Using robotic technology, the resulting culture is distributed into the wells of microtiter dishes and the small cultures in each well are allowed to grow. Each well contains a small-scale culture of C1 containing, on average, a single cloned gene. This collection of micro-scale cultures of C1, each containing a single cloned gene, is referred to as a gene expression library. Robots can be further used to create replicates, or copies of the libraries, which can be stored away for later use.

Libraries are screened – Gene Discovery
Assays for the target, or protein, of interest are developed and the thousands clones in the library are screened for that target. This is accomplished using the C1 High-Throughput Robotic Screening System. Using this robotic screening system, Dyadic estimates it will be able to screen a library of 100,000 clones – enough to cover the entire genome of a simple eukaryote, such as a fungus – in less than one day. Once the activity of interest is discovered, the gene responsible for that activity can be isolated.

Gene Expression
Because the gene discovery process described above targets the product of the gene, that gene is, by definition, expressed at a detectable level during the screening process. However, these levels are usually not high enough to make a commercially viable product. In order to generate enough product for commercial development, Dyadic researches use the "C1 Express" Hyperproducing Protein Expression System to increase the expression level of the gene of interest.

These modern recombinant DNA techniques often result in significantly increased production levels of the protein of interest. Because the same C1 organism is used for gene discovery and gene expression, the probability of successfully increasing the level of protein expression is very high.

Product Development
Once high levels of gene expression are obtained in the laboratory, fermentation development leads to further improvements in the level of product formation. Dyadic’s research engineers and scientists test C1 strains overexpressing the gene of interest by growing them with varying nutrients or under varying environmental conditions. By examining C1 strains in environments with varying temperature, aeration, or pH, for example, the optimal set of fermentation conditions can then be determined.

By choosing the proper conditions, the yield of product from C1 can be improved many fold. In this regard, C1 has advantages over existing expression systems, including other fungi, because of its ability to grow and produce protein under a wide range of growth conditions.

Manufacturing
Once the fermentation process is optimized in the laboratory, scale up enables those processes to become commercial reality. C1 has been in commercial use since 1996, and a number of processes have been readily scaled up from the laboratory to 150,000 liters (40,000 gallons). Yields of protein from C1 are extraordinarily high, making it the system of choice for the economic production of a variety of proteins.

Again, the key advantage of C1’s "one-stop shop" technology is a significantly increased probability of success. By using Dyadic’s proprietary technology, once a useful gene is detected, it can be expressed and has a high likelihood of being overexpressed, to levels that result in a commercially viable product.

By increasing the likelihood of success, Dyadic’s integrated technology has the potential to greatly reduce costs while saving time and resources. By comparison, existing gene expression systems often require the use of a new host organism for product development and production purposes once a desired gene has been discovered and isolated.

Another unique aspect of this technology is the discovery of genes via functional expression in a eukaryotic host using the HTRS system, allowing access to DNA from the entire gene pool. Prior to the innovation of Dyadic’s integrated technology, it was virtually impossible to rapidly screen for useful genes found in the complex life forms that make up more than 90% of the earth’s known species (More at Biodiversity Access).