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UToronto-Spawned Marksman Cellject Pushing Zebrafish Microinjector into Cell, IVF Markets

Marksman Cellject, a Toronto-based company formed to commercialize automated zebrafish embryo microinjection technology developed by a University of Toronto researcher, received a boost last week with the publication of a paper in PLoS One trumpeting a nearly 100 percent success rate for its platform.
The company is now attempting to secure CA$750,000 ($748,000) in seed funding so it can deliver prototypes of its instrument to Canadian and US drug screening labs by the end of the year, company representatives said last week.
In addition, Marksman has partnered with the Toronto Center for Advanced Reproductive Technology and Nikon Canada to parlay a recent CA$100,000 grant from the Ontario Centers of Excellence into further developing the technology for in vitro fertilization and, eventually, for mouse and human genetic applications, the company said.
Marksman Cellject was spawned out of research conducted in the laboratory of Yu Sun, a professor of mechanical and industrial engineering at the University of Toronto.
By training, Yu’s expertise is in micro- and nanosystems mechanical engineering. But Yu, who holds a joint appointment in the UT Institute of Biomaterials and Biomedical Engineering, felt that his work could be effectively applied to increase the efficiency of microinjection of biological specimens, according to Sam Visaisouk, a professional acquaintance of Yu who is currently heading Marksman.
“Sun is a very gifted microrobotics mechanical engineer, and we decided that no one is doing his kind of work in biological materials handling,” said Visaisouk.
Visaisouk was formerly a commercialization manager in the UT Innovations Group, the tech transfer arm for the University of Toronto. Yu presented his technology to the Innovations Group in early 2006, and Visaisouk was assigned to his case.
With the help of Visaisouk and the Innovations Group, Yu won an Idea 2 Innovation grant worth approximately CA$125,000 from Canada’s National Science and Engineering Research Council. Yu used that grant to develop his technology into a fully automated platform for injecting biological materials into zebrafish embryos.
In Canada, assignment of ownership of university-spawned intellectual property varies from university to university. At the University of Toronto, such IP belonged to the researcher at the time Yu and Visaisouk filed US patent applications in mid-2006, though that has since changed. The applications have not yet been published in the US Patent and Trademark Office online database. Marksman has also since filed for a Patent Cooperation Treaty patent application.
As a result of UT’s IP policy, Yu and two of his students own the US microinjection technology patent applications, and will directly receive any royalties from future sales of products related to the technologies, with an undisclosed percentage going to the University of Toronto.
“When the prototype was developed, I saw a business opportunity based on the zebrafish platform to go into other areas,” Visaisouk said. "We talked to the clinic in Toronto, and to some genomics people, and we saw a path to [several] products.”
Visaisouk subsequently incorporated Marksman in September 2006, with Yu as the sole shareholder. “I assist him in negotiations, patenting, and everything other than the research,” Visaisouk explained. “I am not a shareholder. In the future this might change.”
Yu’s microrobotic system incorporates computer vision, motion control, micropipetting, and a specialized array-like embryo holder to automate the microinjection process.
According to the PLoS One paper, published Sept. 12 and on which Yu is lead author, the microrobotic system can inject foreign material such as DNA, RNA, sperm, protein, or drug compounds into zebrafish embryos at a speed of 15 embryos per minute, with a survival rate of 98 percent and a success rate of 99 percent — a far greater efficiency than most available automated techniques, and much faster than manual microinjection.
“If you are interested in genetics, transgenic reproduction, or developmental biology, this is a general tool,” Yu told BTW. “We don’t need human operation, and the machine can operate 24 hours and do it faster than a human being and with better results. If you don’t have a tool like this, it’s not the end of the world. You would just spend a lot of effort doing this, and use graduate students.”
Visaisouk said that another reason Yu started with zebrafish was “because there were a sufficient number of zebrafish researchers on [the University of Toronto] campus to provide him with embryos; and they’re larger [than cells], so it’s a much easier starting point.”
Furthermore, the use of zebrafish in drug screening and genetic analyses has gained in popularity in recent years, with prominent zebrafish programs sprouting up at the University of Oregon, Washington University, and the University of Pennsylvania, among others; as well as with the emergence of biotech companies specializing in zebrafish-based drug screening, such as Zygogen, Phylonix, and DanioLabs, which could provide Marksman with a modest initial market.
But Yu and Visaisouk have designs on expanding the market for Yu’s technology.
“The zebrafish paper was really just one of the things we did — a system we developed to sort of pave our way to the real targets,” Yu said. “Depending on what you’re trying to do — drug discovery, genetics, whatever — if you need a large-scale operation or want better results … you can’t have a human being doing a few samples a day. The machine needs to operate reliably every single minute.”

“The zebrafish paper was really just one of the things we did, a system we developed, to sort of pave our way to the real targets.”

Visaisouk added that “each machine, each model, will be targeted specifically to one application, rather than having a universal machine that’s not really good for everything. So there will be an in vitro fertilization machine, a mouse cell machine, and then a machine for adherent [mammalian] cells.”
Yu is currently developing the IVF instrument with the help of another grant, this one worth about CA$100,000, from the Ontario Centers of Excellence, a Canadian government-sponsored “research-to-commercialization” vehicle; as well as with an undisclosed amount of funding from the Toronto Center for Advanced Reproductive Technology, under the auspices of clinic head and University of Toronto professor of medicine Robert Casper.
Marksman has also scored a partnership with Nikon Canada, which is providing microscopy and certain other mechanical components free of charge with the hopes of landing an OEM contract with Marksman if and when it successfully begins selling its platforms.
“Basically we have access to the latest they have to offer to incorporate into the machine,” Visaisouk said. “They do this free of cost, and we purchase certain components that we will keep. If the relationship develops, that’s no reason for us to have a platform with multiple components; we’ll just stick with Nikon and use their service organization to support our equipment.”
Marksman is not done seeking funding: Visaisouk said that the company is also seeking a grant from the Ontario Genomics Institute to develop an instrument model for mouse cell injections studies. Furthermore, the company is currently attempting to amass some CA$750,000 in seed funding from private investors and a provincial start-up investment fund.
“I hope to close [the funding] before the end of the year,” Visaisouk said. “This will allow us to put a few machines out into the major institutes, and then put it out in the market. I think the US market is where the action is. As soon as we are comfortable that this thing is going to produce the kind of success rate that we set as our objective, we are going to try and get into some very high-profile labs right off the bat.”
Visaisouk said that the initial cost of an instrument for zebrafish embryo injection would cost in the neighborhood of CA$200,000.
Marksman will have to achieve its goals without further assistance from the UT Innovations Group. Visaisouk left his job at UT approximately six weeks ago to help focus more on Marksman’s development. Since then, he has taken a new tech-transfer position at the University of Waterloo in Ontario, but will continue to head Marksman until greater organizational structure is needed.
“The company is very much independent of the university,” Visaisouk said. “For me, I’m just helping him as a friend, because the company has no money. All the money is dedicated to R&D and developing prototypes. Once we put machines out, we will need to develop an infrastructure to support that, and we don’t want to pull away from [Yu’s] research.”

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