Better Know a Scientist: Estefania Elorriaga

This week in “Better Know a Scientist”, I’m interviewing Estefania Elorriaga. She’s in the midst of her PhD in Dr Steven Strauss’ lab in the Department of Forest Ecosystems and Society at Oregon State University. She is doing research on using site-specific nucleases for mutagenesis (fear not! She’ll have to explain her research in this interview).

Like me, Estefania was raised in Venezuela and some of the comments and questions in this interview center around Venezuela’s current political climate, so a bit of a background is needed: Hugo Chávez was elected as Venezuela’s president in 1999 and remained in power till his death in 2013, after which his appointed candidate was elected into office. Chávez led a “socialist revolution” that took a very hard anti-American, “anti-imperialist” stance. As such, much of the political discourse in the country reflects this attitude with frequent conspiracy theories about the so-called “American empire’s” involvement in the country.
Let’s get started!
Q: What are site-specific nucleases? Why are they important?
Site-specific nucleases are enzymes that can cut DNA at specific  locations in the genome of your organism.  The nucleases create a break which stimulates the organism’s DNA repair mechanisms to fix the break.  Occasionally, the repair mechanisms will make a mistake (delete some DNA or insert some extra DNA) that will lead to a loss-of-function mutation in the target gene, meaning that the protein you targeted will no longer do its job.  Examples of site-specific nucleases are zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), the CRISPR (clustered regularly interspaced short palindromic repeats)-Cas system, and engineered meganucleases.  Site-specific nucleases can also be used for gene repair or gene replacement with the addition of a donor DNA sequence.  These technologies are important because they allow scientists to modify only specific locations of genes in order to obtain a desired trait.  In the area of crop breeding we believe these technologies will be crucial for improving current lines specially for particular cases like drought-tolerance, high or low temperature tolerance, disease resistance, or even nutrient content.
[Layla’s note: if you want to learn more about gene editing, please see my previous post on the topic]

The plant on the left has a TALEN (will show it's trait much later) The plant on the top-right is her transgenic control,  expressing GFP (green fluorescent protein). The plant on the bottom right is the unmodified control.
The plant on the left has a TALEN (will show it’s trait much later)
The plant on the top-right is her transgenic control,
expressing GFP (green fluorescent protein).
The plant on the bottom right is the unmodified control.

Q: How specific are the nucleases? Are you seeing off-target edits in your work?
We are still at the early analysis stage, so I cannot really say personally how specific they are.  But, according to the scientific literature, CRISPR-Cas nucleases appear to be the most active and specific, followed by TALENs, and last ZFNs.  CRISPR-Cas nuclease show a lot of promise because of the high activity that they show so far and their off-targeting appears to be somewhat predictable.
Q: Are there any GMOs made using site-specific nucleases currently on the market? Do you know of any that are in development?
There are no commercially available GMOs currently in the market or in development (that I know of), but everyone in the field believes that the technology is going to revolutionize genetic engineering, especially breeding and gene therapy.
Q: Like me, you were raised in Venezuela. Venezuela has a moratorium on growing any GMOs and activists would argue that the moratorium is in place because of how harmful GMOs can be. So, how do we know that you aren’t a spy sent by Venezuela to the United States to bring down the imperialist empire through evil GMOs?
Ha ha! If the Venezuelan government was making me get a PhD just to spy on the USA, I think I would have quit my job as a spy and gone to Colombia or Costa Rica.  I cannot imagine doing research while spying.  As it stands, I barely have any free time.  And also, I cannot imagine pretending to be a researcher.
[Layla’s note: that’s exactly what a person tasked with destroying the imperialist empire with GMOs would do: deny it].
Q: What traits are you working on in the lab? Why are they important?
I work with flowering genes in both poplar and eucalypts. Understanding floral development and identity genes is important because it will allow us to generate transgenic trees that don’t have functional pollen or ovules, so there is no possibility of transgene flow.  Developing transgene containment technologies for forest trees to facilitate the commercial and scientific use of transgenic trees is one of the main goals of our lab.

Q: Can you explain what you mean by “transgene flow”? [Layla’s note: some activists refer to this as “GMO contamination”] Does it basically mean that you’re blocking the GM tree or plant from hybridizing or crossing with a non-GM tree or plant?

“Transgene flow” is the movement of the gene we inserted into our trees into a wild or cultivated population of the same or a related species. And yes, it basically means not allowing our GM tree to cross or hybridise with wild or cultivated trees.

Q: If you’re creating a GMO to address one of the major concerns against GMOs, aren’t you creating a circular argument? How would that conversation even go? I imagine it would be like, “Activists! You’re worried about GMO genes getting into the environment? Here’s a GMO that will prevent GM genes from getting into the environment. Go forth and plant it!” Aren’t a few heads going to explode? Is THAT the real plan of the Venezuelan government? To bring down the imperialist empire through circular arguments?

By eliminating transgene flow, we are just allowing the GM trees to be used commercially for other purposes like a non-flowering faster growing eucalyptus.  Also, eliminating transgene flow in the current GM crops will allow the scientific community to educate the public, and also other scientists that are weary, about the safety of GM technology.  We hypothesize that transgenes will likely not do well in the wild, so they will probably get eliminated through fitness selection, but we need to study each particular case. Ha ha! Nothing from the current Venezuelan government surprises me, so who knows…
Q: Are there any non-GM crops where gene-flow is a problem that may benefit from the adoption of the technology?
The other case that comes to my mind is the case of gene purity in seed crops.  Seed producers follow standards and practices that guarantee the purity of the seeds available to farmers and home growers. Genetic purity in their case is essential to ensure that the seed will perform as expected, so seed producers must be careful about the pollen that fertilizes their plants.
Q: What trait would you like to work on in the future?
I would like to work with either nutrients (e.g. create a highly nutritious fruit or vegetable crop), drugs (e.g. create plants that can make medicines), or abiotic resistance (e.g. create plants that can take up heavy metals).
Q: There are many articles in the news and in journals about potential traits that may benefit us, many of which never make it past research, but none-the-less create much buzz. Which one(s) do you think are the most exciting? Which one(s) do you think are the most promising?

Estefania Elorriaga
Estefania Elorriaga

Being a transplant in Oregon and feeling like a true Oregonian since I moved to the Northwest, protecting the environment is a topic dear to my heart.  In the University of Washington, Prof. Sharon Doty worked with transgenic poplars that were able to remove more than 90% of contaminants from Superfund sites. According to the EPA, “A Superfund site is an uncontrolled or abandoned place where hazardous waste is located, possibly affecting local ecosystems or people”. I think using trees to clean our aquifers and our soils is a win-win. We get more oxygen, cleaner air, and also cleaner soils and rivers. But, the trees are nowhere near being ready for use outside of university labs.
There is also a transgenic pig called “the Enviropig” from the University of Guelph in Canada, that digests phosphorus from its food more efficiently than non-transgenic pigs, so it needs less feed and its waste is less toxic to the environment.  Doesn’t that sound like a win-win too?!
I am also a big fan of producing medicines in plants (like I mentioned above).  This practice is known as biopharming.  I am also a fan of using GM mosquitoes to eliminate vector-borne diseases like dengue (after the GM mosquito mates with a non-GM mosquito the transgene causes the mosquito larvae to die).  These two cases I mentioned though are closer to production than the Enviropig or phytoremediating trees.  Biopharming had a rough time recently because many of the companies involved went bankrupt, but there is an experimental antibody for Ebola currently being made in tobacco by Kentucky BioProcessing called ZMapp that should give this industry a boost, and the British company Oxitec plans to release GM mosquitoes in the Florida keys to reduce dengue and chikungunya (both diseases you don’t want to get, and less yet while on vacation).  ZMapp is considered the most promising candidate to combat Ebola and it is currently being used in a controlled human trial in Liberia. The mosquitoes were already tested in Brazil and Panama with great success, so both countries plan on releasing a lot more of them.
[Layla’s note: if you want to learn more about Oxitec’s mosquitoes, please see this post on my family’s experience with dengue and my review of the literature on these phenomenal skeeters]
Q: How much money is Monsanto paying you to develop these transgene containment technologies that will solve one of their public image problems? After all, we all know that Monsanto controls university research.
Monsanto has never paid me or our lab for the research we do.  Sadly, when many people think “GM technology” they think “Monsanto”.  But, in reality the technology is being used by hundreds of university labs, research facilities, and biotech companies developing more than herbicide- or insect-resistant crops.  Monsanto and all the other large biotech multinationals like Dupont, Pioneer, or Syngenta come up with cutting-edge seed and agro-chemical technologies, but given that there is no direct benefit to the end user (all the benefits go to the farmer), the public shows little to no support because of lack of understanding and fear.  If you look over the ISAAA’s (International Service for the Acquisition of Agri-biotech Application) Pocket K. Documented Benefits of GM Crops, you will find that from 1996 to 2012, global farmers’ income increased by $116.6 billion and there was a reduction in herbicide and pesticide use of 503 million kgs (ISAAA is an international non-profit “that shares the benefits of crop biotechnology to various stakeholders, particularly resource-poor farmers in developing countries, through knowledge sharing initiatives and the transfer and delivery of proprietary biotechnology applications”).  All these agricultural biotech companies perform cutting edge science and create impressive biotech products.  However, as companies they are concerned with profits, so given the high cost of generating a GM product, they focus on products that will bring them a return on their investment.
Q: My dreams for crop modification aren’t as lofty: I just want a peelable pomegranate. Do you think that the research you’re conducting might be able to help?
My work won’t directly affect the possibility of creating a more peelable pomegranate. But, indirectly it can bring science closer to your dream.  My research can add onto the increasing scientific knowledge base that is proving that site-specific nucleases can someday be an important tool in crop breeding.  If we find the gene or sets of genes involved in making the pomegranate’s skin, we can either modify or replace the genes for ones that will make the pomegranate easier to peel.  For many centuries, humans have been modifying the aspect, size, and taste of cultivated crops by doing selective breeding without knowing anything about genetics.  Wild bananas and corn (and dogs…chihuahuas come from wolves!) are great examples of what selective breeding can do.  Wild bananas are small and loaded with seeds.  Meanwhile, cultivated bananas are large and have tiny seeds (that actually are not viable because cultivated bananas are sterile).  Teosinte (the wild ancestor of corn) make small ears with only two rows of hard fruit cases that protect the seeds.  Corn makes large ears with eight to twelve rows of tender seeds (no hard fruit cases).  So, GE is basically streamlining breeding by allowing use scientists and plant breeders to perform highly selective and direct breeding using genes from the same species or other species.
[Layla’s note to her husband: it sounds like there’s the remote possibility that Estefania’s research might be able to make my pomegranate. Please be advised that we may have to move to Oregon in the near future to help conduct this research].


  1. “abiotic resistance (e.g. create plants that can take up heavy metals)”
    Isn’t that phytoremediation? Abiotic resistance is, at least to my understanding, drought tolerance, flood tolerance or capacity to grow under limiting nutrient conditions (and assorted others). /nitpick
    One other very important aspect of site specific nucleases, and the development of this technology, is that it will enable the sort of gene stacking that will be required to further agricultural use of GMOs. At present we’re at a maximum of about 8 genes (whatever it is that smartstax is) which must all be tracked during trait introgression when developing new varieties. This is a costly and time consuming process, and quite frankly at present likely represents one of the biggest obstacles to increasing the number of stacked genes.
    If transgenes could be targetted, accurately, within the genome, one could essentially simplify the process back down to a regular single trait introgression rather than a complex multi-trait introgression – each transgene would be inserted next to (or close as is meaningless anyway) the other transgenes you wished to keep together and you’d remove the time/cost barrier to new trait introduction (which is relatively hidden, but a big deal – consider that ~100 corn hybrids are released per year in the US by Monsanto per year, with new lines constantly being added to the mix – these lines all need to have traits introgressed into them in order to be viable, and when dealing with something like smartstax this process is a tremendous challenge.

  2. Ewan,
    What about promoters? In practice, do these transgenes come integrated each with an appropriate promoter, or do they have to be inserted in specific places to “share” existing ones?

  3. Hi Ewan,
    Yes, the use of plants to remove contaminants is indeed phytoremediation. Nonetheless, phytoremediation with wild type plants is slow and not effective in places that have high concentrations of contaminants. Thus, the transgenic poplar are considered “abiotic stress resistant” because they can tolerate and metabolize contaminants more effectively.

  4. Hi orchidgrowingman,
    In practice, transgenes are generally integrated with the appropriate promoter (constitutive, tissue-specific, developmental stage-specific, etc.). That said, site-directed nucleases can be used to insert a cis/transgene in a specific location (e.g. next to a specific endogenous promoter) by having a donor sequence (i.e. the gene to be inserted) close by when the repair enzymes are working on the cleavage site.
    Also (not specifically related to your question but interesting), there is a technique known as “promoter trapping” where hundreds to thousands of transgenic events are generated with a promoter-less reporter gene (like green fluorescent protein from jellyfish) with the purpose of identifying tissue- or developmental stage-specific promoters.

  5. I imagine if one was wholesale replacing a gene (with similar technology, it’d get a little more complex) then one could target to a plant specific promoter – but the genes I’m thinking of all have their own promoter and terminator (not that kind of terminator…) regions – they’d also likely need some buffer around them in order to prevent promoters interfering with each other etc.
    One would assume that one of the main priorities for a region to insert genes (particularly a bunch of them) would be a region that is pretty much devoid of any genes while, at the same time, has been shown to be transcriptionally active (as it’d really suck to blow all your resources shuffling genes into a region that was silenced all the time (by whatever means))

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