CRISPR 101: Applications and Future Potential
By Kevin Holden, PhD, Head of Synthetic Biology at Synthego
This is the second part in the CRISPR 101 series by Synthego, providing a crash course on CRISPR-Cas9 and its applications in a wide range of life science disciplines.
Due to its versatility, CRISPR-Cas9 genome editing technologies have the potential to accelerate synthetic biology research and development across a variety of industries. Given that the world’s population is expected to reach close to 10 billion by 2050, such advances are welcome as we look to discover new disease therapies and feed a burgeoning population. CRISPR genome engineering can help medicine to become more personalized for the patient, and will also aid researchers in reacting quickly to emerging epidemics. In a future of shifting climate extremes and migratory populations, the ability to increase yields and engineer adaptations to pests, drought and disease in our food sources will become increasingly important. Furthermore, the ability to quickly bioengineer organisms for biofuel and textile production will help the world address diminishing bioresources.
How can CRISPR-Cas9 genome editing have a positive effect on such a wide range of applications? Read on to find out how it is already having an impact toward a number of research and development activities across many disciplines.
CRISPR has exponentially increased how quickly and efficiently the genomes of immortalized cell lines can be engineered. Although other gene editing technologies existed before CRISPR, such as Zinc Finger Nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs), these were typically less efficient, limited by the number of specific locations within a genome that they could target, expensive and time-consuming to generate and utilize. CRISPR, however, requires only two or three components (Cas9 nuclease, guide RNA and optional donor DNA) that are readily available from many sources at practical price points, scales and delivery times. Furthermore, CRISPR can be performed in most existing molecular and cell biology laboratories without the need to invest in expensive new equipment. Using CRISPR, the process of generating gene knockouts, knock-ins and single nucleotide variants (SNVs) in cells in order to model disease has become significantly faster and orders of magnitude more efficient. As more labs, for example, are able to generate cancer genotypes in specific cell lines, more potential targets can be identified and more pharmaceutical drugs evaluated – speeding up the process by which we identify new cures for cancer.
Another critical component to studying infectious pathogenesis, autoimmunity, cancer, mechanisms of action of pharmaceutical therapies and drug discovery in general is the ability to generate transgenic animals to model disease. Mice are the most model for many of these applications. Before CRISPR, generating transgenic mice for these applications could be time-consuming and inefficient – sometimes taking up to a year to produce a correct genotype. Much like with cell lines, CRISPR can be used to generate gene knockouts, knock-ins and single nucleotide variants (SNVs) in mouse embryos and this has resulted in a dramatic increase in how fast transgenic mice can be generated. In addition, the increased efficiency at which they are generated results in less overall mice being used in this process.
Personalized Medicine & Gene Therapy
The promise of gene therapy has remained unrealized for some time now. However, with the emergence of CRISPR, the future for such personalized medicines looks bright. While the first published report of an inherited disease mutation corrected in a human embryo (that was allowed to divide to blastomere stage) has generated some early hope for the genetic treatment of rare genetic disorders, less complex gene therapies are currently entering clinical trial phases and could become a reality in the near future. This includes the ability to utilize CRISPR to edit a patient’s own stem cells to repair, for example, a blood disorder such as Sickle Cell Anemia, and then re-implant and expanded number of these edited stem cells back into the patient in order to correct the pathology. Beyond genetic disorders, this process can also be utilized by editing a patient’s stem cells to generate T-cells that are resistant to HIV infection. Another exciting treatment involving T-cells that CRISPR is helping to accelerate is the development of chimeric antigen receptor T-cells, or CAR-T therapies. In this case, a patient’s own T-cells – the orchestrator of many immune responses – are isolated and edited in the laboratory in order to target them towards that patient’s cancer.
Perhaps a more far-reaching application of CRISPR with respect to the world’s growing population, beyond disease therapies, is the generation of transgenic plants. In order to satisfy increasing food demands and to enable farming of crops in areas of the world that were once thought unworkable, advanced genetic breeding techniques will need to be employed. In addition, changing weather patterns could introduce droughts, freezing or new pests into existing agricultural zones. Traditional methods for breeding plants and animals used for food, although time-tested and present in most of the foods we consume today, are very slow – sometimes taking generations to develop, and are not amenable to change. With CRISPR, the time at which it takes to edit plant genomes has increased significantly. While it could previously take several years to generate new varieties of crops, either using traditional breeding or genetics, such diversity generation can now be completed in a matter of months. Already, CRISPR is being used to breed plants that are more resistant to drought, disease and pests.
Biofuels & Specialty Chemicals
Our growing population is also leading to a reduction of non-renewable bioresources, such as coal and crude oil. Compounding this problem are the environmentally destructive nature of many next-generation mining techniques, such as fracking. Synthetic biology is providing solutions for our reliance on petrochemicals and their derivatives by utilizing CRISPR to engineer microbes and plants to generate biofuels and specialty chemicals. These compounds can be used to generate the plastics, textiles, fragrances, lubricants and fuels that millions of people use each day. Engineering microbes and plants to operate as chemical factories in large-scale fermentation processes has traditionally been challenging. With CRISPR, the numerous genetic manipulations required to complete this bioengineering in yeast, bacteria and algae can now be performed at a much faster rate than previously possible. This in turn yields to an accelerated rate at which novel strains can be screened for chemical production and composition before being evaluated at the large-scale fermentation stage. Already, CRISPR is being utilized to engineer bacteria, yeast and algae for the production of hydrocarbons – the precursors for many fuels and specialty biochemicals – and for alternative fuels such as ethanol.
Synthego is a leading provider of genome engineering solutions. The company’s product portfolio includes software and synthetic RNA kits designed for CRISPR genome editing and research. With next-generation informatics and machine learning, Synthego’s vision is to bring precision and automation to genome engineering, enabling rapid and cost-effective research with consistent results for every scientist.
Headquartered in Silicon Valley, California, Synthego customers include leading institutions in over 32 countries around the world, 8 of the world’s 10 largest biotechnology companies, and 24 of the top 25 global biology universities.