by Jestin George
In 2012 the first evidence of CRISPR-Cas9 genome editing was published and since then, almost 4000 CRISPR-Cas9 articles have followed. From fundamental research to applications in biotechnology, agriculture, health care, pest control, and synthetic biology, there is no doubt that CRISPR systems will revolutionise both biological research and indeed, technology, as we know it. The astounding rate of progress over the past 5 years has not been ignored: policies are being revised and reconsidered, moratoriums and summits are being held globally, all to ensure the progression of safe and ethically driven genome-editing technology.
Recently, two jam-packed genome-editing conferences took place in Australia, both focusing on socially relevant areas of research: cancer treatment and agriculture. These meetings were aptly timed given the political landscape concerning gene-editing technology. The National Institutes of Health, USA approved CRISPR-Cas9 as a therapeutic for cancer and the Australian Office of Gene Technology Regulator (OGTR) is undergoing a technical review of its regulations. It’s Gene Technology Regulations Act (2001) is to be amended to provide clarity about how Australian law defines genetically modified organisms (GMOs). This is important because current legislation has not kept up with advancing technologies, resulting in some strange double standards. For example, two identical organisms can be defined differently (GMO and non-GMO) because of the process used to generate them. Using a simple “process-based” and not “product-based” definition of a GMO is now inappropriate and outdated. This technical review aims to implement a more evidence-based policy for assessing risk and safety. The OGTR is consulting the public on the matter until February 2018; don’t miss the opportunity to have your say here.
The Genome Engineering for Cancer Treatment meeting (The Frank and Bobbie Fenner Conference) was held at Australian National University (ANU). One week later came the CRISPR-Cas9 in Cereal Crops workshop held by CSIRO in the beautiful seaside town of Kiama. While both meetings served as opportunities to hear about cutting-edge research in these fields, at the core these meetings showcased where the research in Australia is. Consequently both meetings provided the chance for a great deal of knowledge sharing, particularly regarding technical difficulties in applying the technique. They also both provided important non-scientific information, including: the patent and licensing landscape of CRISPR systems, bio-ethical and social considerations and strategies, and information regarding the OGTR’s technical review.
Australian Chief Scientist Dr Alan Finkel opened the Cancer Treatment meeting entitled “Excellence with Equity and Empathy”. In his speech, Dr Finkel offered three mantras for the success of scientific agendas: (1) speak human to humans (2) think out to the ten-year horizon and (3) regulate to facilitate, which resonated through the talks given. Dr Gaetan Burgio presented some sobering evidence for poor reproducibility in the field, attributing it to low sample sizes, lack of lab-based skills and scant reporting of methodologies. There was also a spotlight on CRISPR bioinformatic and biostatistic research. Keynote speaker Dr Veera Baladandayuthapani unpacked their work in discovering relevant targets using biostatistics. Dr Laurence Wilson discussed machine-learning approaches for improving CRISPR targeting efficiency and introduced their optimised target identification platform, GT-Scan2. This software incorporates a range of previously overlooked factors that affect editing efficiency, including chromatin structure and epigenetic markers.
Epigenetic editing itself was covered by Prof. Ryan Lister, who demonstrated that for heritable reconfiguration of chromatin by editing, simultaneous activity of multiple, distinct epigenetic effectors is needed. Assoc. Prof. Pilar Blancafort showed how they used epigenetic editing for reprogramming complex phenotypes in breast cancer. Dr Ben Hayley compared the various cutting-edge CRISPR activator (CRISPRa) systems available and presented the PiggyBac transposon for large cargo transgenesis, size being a limiting factor regarding viral delivery systems. But it was the extent of genome-wide editing applications being used in cancer research that was powerful to see. Such applications presented included: screening novel tumour suppressor genes, unpacking the hidden details of the critical tumor suppressor gene p53 and JAK-STAT signalling, identifying resistance mechanisms in multiple myeloma, and identifying novel drug targets for generating functional organoids in colorectal research. As Dr Finkel mentioned in his speech, Australia will be a player in the age of genome medicine.
The CRISPR-Cas9 in Cereal Crops meeting opened with an impressive overview of metabolic engineering by CSIRO’s Agriculture and Food innovation leader Dr Allan Green. It was inspiring and motivating as a biotechnologist to hear about the success of Australian companies GoResources and NuSeed -which use engineered organisms to provide clean energy and improved crop productivity- as well as NxtOil technology. Prof. Caixia Gao showed their impressive work on wheat editing, and is soon to publish a novel protocol for biolistic delivery of Cas9 ribonucleoprotein (RNP). Prof. Gao also demonstrated an in vivo screen for assessing optimal CRISPR guide RNAs (gRNA) using protoplasts instead of in vitro DNA digests. Given the research presented by Dr Wilson regarding the effect of chromatin structure and epigenetic markers on Cas9 targeting efficiency, this in vivo validation technique is an important tool for plant editing. Yao-Guang Liu presented their work on DSDecodeM, an online tool for automating chromatogram analysis. During screening, chromatogram analysis usually requires manual analysis of sometimes-vast numbers of sequencing reads which can be very time-costly.
Prof. Jimmy Botella gave a hilarious but impressive overview of their CRISPR-mediated ultrafast breeding for adding one or two traits to an otherwise elite line in just two generations. Dr Mike Jones demonstrated how they used gene editing to create low gluten index (GI) potatoes. He also spoke to the importance of genome editing techniques for modifying properties of existing cultivars without introducing external DNA, given the lengthy time frame of breeding and market acceptance. Dr Mick Ayliffe commented on the benefits of gene stacking for durable, longer-lived pathogen resistance by delivering numerous genes on one locus. He also showed their unsuccessful efforts to bring a “pseudogene back from the dead,” an attempt to restore resistance traits. It served as a reminder that destroying gene function is much easier than creating it, even with gene editing tools like CRISPR.
Dr Heather Bray, Dr Lucy Carter, Dr Aditi Mankad and Craig Cormick all gave fantastic insights into dealing with social acceptance and considering the broader implications and ethics. They reminded scientists to also think like human beings, echoing Dr Finkel’s opening speech. Food is not only about sustenance; it is about ceremony, ritual. Values, not information, determine views. People need to know who is benefiting from what technologies, both directly and financially, more than they need to understand the science. While the societal and ethical elements to CRISPR are tricky to navigate, one thing that is clear is that the deficit model –the idea that the public would be more on board if only they knew as much as us scientists– does not work.
While evidence of the widespread success and application of CRISPR technology is clear, the CRISPR revolution is only just beginning. We are excited that Australia, as Dr Finkel commented in his speech, is going to be a key player.
Jestin George is a PhD student at the University of Technology Sydney, working on genetically engineering the microalga, Chlamydomonas reinhardtii, for applications in biotechnology. She is member of the executive board and assistant communications officer of Synthetic Biology Australasia.