BORNHEIM, Germany - The German company Mermaid Pharmaceuticals GmbH and U.S.-based Gene Tools LLC entered an exclusive research alliance in zebrafish functional genomics.

The companies plan to functionally analyze the majority of vertebrate genes at the rate of hundreds of genes per week, Mermaid's president, Alexander Crawford, told BioWorld International. Financial details of the deal were not disclosed.

"The zebrafish is an excellent model of human biology for the rapid identification and validation of disease-relevant drug targets," Crawford said. "Currently, most vertebrate genes are known by their sequences but not by their functions. Mermaid will discover the functions for many of these genes by high-throughput reverse genetics in the zebrafish."

For this, the Hamburg-based company will use Corvallis, Oregon-based Gene Tools' Morpholino technology. Morpholino oligomers are antisense molecules that bind and inactivate selected RNA sequences, according to Gene Tools.

"Once we know the sequence of a gene, we can quickly design a Morpholino oligomer that blocks that gene's translation. We don't even need the entire sequence of the gene," Crawford said. "We inject Morpholino oligomers into early stage zebrafish embryos and observe in the hours and days that follow the biological effects of the inactivation of these genes. We don't need to wait for the results of time-intensive breeding schemes and gene-cloning procedures necessary in classical forward genetics approaches."

If the Morpholino blocks translation of a gene necessary for blood differentiation, for example, there might be too little or no blood, and this can easily be detected in the zebrafish embryo. If the oligomer inactivates a gene involved in cartilage formation, then there may be too much or too little cartilage in the embryo afterward.

"We quickly detect such changes through a battery of molecular and phenotypic assays, including simple light microscopy, within the first few days after injecting the Morpholinos," said Crawford, who added that that Mermaid scientists also will be analyzing the expression of specific pathway markers by semiautomated detection of fluorescent reporter proteins in zebrafish embryos maintained in 96- and 384-well plates.

"We consider the screening of hundreds of genes per week by these methods to be true high-throughput target validation," Crawford said.

"Mermaid will generate revenue by discovering and patenting drug targets with which to enter alliances with pharmaceutical companies," Crawford said, adding that his company also can help big pharma validate and prioritize existing candidate targets for drug discovery. "The company is currently in discussions with several potential pharmaceutical partners," he said. "But at the moment we are not releasing further details."

Crawford was a founding executive of Ingenium Pharmaceuticals AG, a Munich, Germany-based mouse genetics firm. He left that company in 1999, he said, to pursue zebrafish-based technologies, founding Mermaid together with Camila Esguerra, Michael Ratigan and Dirk Sebastian. Mermaid initially was funded by its founders.

It disclosed its first major financing round on Nov. 2. The company raised EUR13 million (US$11.5 million) through German investors BioAgency AG, of Hamburg; Haspa BGM, of Hamburg; TFG Venture Capital, of Marl, Germany; and 2Venture AG and tbg, both of Bonn.