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Interview with Dr. Keith Pardee
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We read your paper from last year titled “Rapid, Low-Cost Detection of Zika Virus Using
Programmable Biomolecular Components” and we wanted to know, what led you to
use β-galactosidase as a colored readout?
Dr. Pardee: At that time, we hadn’t considered using C2C2 (Cas13a). So, one of the reasons was
because of the enzyme activity. Single molecule of reporter could serve amplification, rather than
just fluorescence. The other (reason) was just for practical use. No need (to use) UV lights,
electronics, camera, and could be done in a simple piece of paper. Tried 10 different enzyme
options with other based reporters.
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What was your motivation to build that detector on your study?
Dr. Pardee: Multiplexing, being able to track multiple reactions at the same time, and the
potential for quantification. These processes are semi-quantitative and so with that information,
you want to be able to read rather than just being plus minus, being able to calibrate your
reactions. You might be not being able to say exactly how much you have but you may be able to
say this is a high titer or a low titer individual. Also, user accessibility, if you can automatize some
of this tests for reading like in pregnancy tests with a digital detector… can make it really easier
to use.
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Regarding our project
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What do you think about the problem of antibiotic resistance and the ways to prevent
it?
Dr. Pardee: This is probably the biggest question and it is not my area of expertise. We should
continue the advocacy of responsible use in agriculture and in patients. With tools that we both
are building where we can identify resistance early and contain it and treat individuals that have
resistance is also going to be important.
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We have some problems getting the Cas13a lyophilized into paper. Do you have any
low-cost suggestion for us?
Dr. Pardee: I have never done that but there was a new paper published (Kikuta et al., 2017) that
uses trehalose as a cryoprotectant, that lets you dry cell free-reactions at room temperature. The
trehalose is present in water bears and seeds, and helps keeping things dry. It is an inhibitor of
cell free-reactions at high concentrations.
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Did you ever considered using as a colorimetric readout such as gold nanoparticles?
Dr. Pardee: Yes absolutely. I think is a great thing to do and useful. You can borrow so much from
what is already been done.
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Regarding the Synthetic Biology field
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What difference can Synthetic Biology bring to the world? How significant or relevant
can it be for future generations?
Dr. Pardee: As synthetic biologists we have a real responsibility to be transparent and to
communicate with the public actively, not just waiting for the public to come to us but we need
to see that the public knows what we are doing why we are doing it. Especially because there are
real tensual concerns and also perceived concerns. I think being ahead of those and managing
them responsibly is important. Otherwise there will be a repeat of what happened with
genetically modified plants in the 80s-90s.
I think Synthetic Biology really has tons of potential to solve a lot of the big challenges of the day
and I think probably the biggest one is inequity. We are very fortunate in the parts the world
where we live, we have access to high quality food, sufficient, quality health care and energy and
large parts of the world don’t have. I think that synthetic biology and biological engineering and
science in general can sort of rate /change this inequality by creating tools that are low cost and
sort of spread that the wealth that is in present in one part of the world, not in others to any
other part world. I think synthetic biology can address energy food, and health. The work that we
are doing in our lab addresses some of those aspects. The fact that our work and your work is
cell free is also trying to address concerns of biosafety. Cell-based technology can also be safe if
implemented in responsible ways. In our case, we are working in diagnostics and access to drugs
and there are million other ways to use Synthetic Biology. The nice part from a scientist
perspective, it´s (that we are in the) early days and the space is wide open, so whatever drives
you and wherever your creativity takes you. That´s exciting.
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"As synthetic biologists we have a real responsibility to be transparent and to
communicate with the public actively, not just waiting for the public to come to us but we need
to see that the public knows what we are doing why we are doing it."
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What was your motivation to build that detector on your study?
Dr. Pardee: Multiplexing, being able to track multiple reactions at the same time, and the
potential for quantification. These processes are semi-quantitative and so with that information,
you want to be able to read rather than just being plus minus, being able to calibrate your
reactions. You might be not being able to say exactly how much you have but you may be able to
say this is a high titer or a low titer individual. Also, user accessibility, if you can automatize some
of this tests for reading like in pregnancy tests with a digital detector… can make it really easier
to use.
|
Regarding our project
|
|
What do you think about the problem of antibiotic resistance and the ways to prevent
it?
Dr. Pardee: This is probably the biggest question and it is not my area of expertise. We should
continue the advocacy of responsible use in agriculture and in patients. With tools that we both
are building where we can identify resistance early and contain it and treat individuals that have
resistance is also going to be important.
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