The announcement of being able to achieve a higher than 14-15 percent transfection efficiency with low mortality in motor neurons is very exciting news—not just for ALS researchers, but for anyone studying motor neuron-based biology. One of our issues with doing studies in primary mouse motor neurons is having a high enough transfection efficiency, with sufficient viability to conduct meaningful studies with more than just a handful of transfected motor neurons.

The methodology described by these authors would be easier to implement in a general lab where there are safety concerns and special protocols that must be followed when working with lentiviral infection, which is the current system for moving transgenes to these cells. This methodology does not employ an infectious agent. This is extremely attractive, at least in Canada, where very specific regulations and protocols for lentivirus use exist.

That the authors can use this with silencing experiments as well is encouraging. Knockdown levels do not seem to be very high, a common problem with silencing in many cell...  Read more

The announcement of being able to achieve a higher than 14-15 percent transfection efficiency with low mortality in motor neurons is very exciting news—not just for ALS researchers, but for anyone studying motor neuron-based biology. One of our issues with doing studies in primary mouse motor neurons is having a high enough transfection efficiency, with sufficient viability to conduct meaningful studies with more than just a handful of transfected motor neurons.

The methodology described by these authors would be easier to implement in a general lab where there are safety concerns and special protocols that must be followed when working with lentiviral infection, which is the current system for moving transgenes to these cells. This methodology does not employ an infectious agent. This is extremely attractive, at least in Canada, where very specific regulations and protocols for lentivirus use exist.

That the authors can use this with silencing experiments as well is encouraging. Knockdown levels do not seem to be very high, a common problem with silencing in many cell types. Still, this methodology should help with studies in overexpression of proteins by knocking them down again to more physiological levels, or even taking a non-transfected expressed gene and knocking that down to determine what effect that has on the neuron.

I hope that the authors will follow up with a determination of expression levels of proteins in this system. It would be useful to see just how overexpressed a transfected construct is or can be, and if those levels can be manipulated with various promoter systems (as can be done in other cell types) to force expression of transgenes to disease state appropriate levels. It would also be very interesting to determine if this system could be efficient for the transfection of adult-derived mouse motor neurons, and what the viability courses of both day 13.5 and adult-derived motor neurons are post-transfection for longer-term studies.

View all comments by Kathryn Volkening