Genetic Engineering & Biotechnology News

NOV1 2018

Genetic Engineering & Biotechnology News (GEN) is the world's most widely read biotech publication. It provides the R&D community with critical information on the tools, technologies, and trends that drive the biotech industry.

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Page 16 of 57 | Genetic Engineering & Biotechnology News | NOVEMBER 1, 2018 | 15 OMICS technique must become less complex, and this will happen by making instruments smarter and preconfiguring reagents. Ms. Talaga: Since its inception, flow cytometry has been important in clinical diagnosis. More re- cently, flow cytometry has been used to im- prove the analysis of small particles such as circulating exosomes, which promise to in- form liquid biopsies. Now, flow cytometry is heading into single-cell analysis. As flow cytometers become more capa- ble—pushing the limits of color detection, for example—they allow us to collect vast amounts of information from small sam- ples. Small vesicles have become important because of their biological significance, but instead of trying a number of other tech- niques, we can now turn to flow to simplify this area. We expect that in the near future, more scientists will either conduct analysis of single cells, in both biological and clinical samples, or verify processes by flow. GEN: How is flow cytometry contributing to the "CRISPR craze" currently rocking the life sciences? Dr. Guenther: CRISPR technology has a strong potential for the treatment of various diseases, either directly or indirectly. As a CRISPR tool, flow cytometry is important not only for validat- ing the correct targeting performed by CRIS- PR, but also for measuring the functional effects of gene editing. For example, flow cytometry can be used to screen cells for proper transfection, allow- ing for rapid high-throughput analysis of many different reactions. It can also be used to identify functional targets for CRISPR, es- pecially in immune cells to monitor cytokine production and changes in surface markers. Additionally, the flow cytometer can easily be utilized as a screening platform for CRIS- PR, especially with the advancement of high- throughput autosampling capabilities. Ms. Wright: As genome editing techniques become rou- tine, demand is increasing for cell analysis tools. At the simplest level, if we modify the "programming" of a cell, we impact its func- tion, so we need techniques to help us ana- lyze what happens downstream. Have we disrupted, induced, or modified protein pro- duction? What is the efficiency of the modifi- cation within a target cell population? Have our changes affected other aspects of cellular function? Are there unexpected or off-target effects? Flow cytometry and functional cell sorting will be increasingly important tech- niques to help answer these questions and more as the field of genome editing evolves. Ms. Talaga: In the past few years, we have seen an explo- sion of companies utilizing flow cytometry in the CRISPR process, doing everything from determining transfection rates to sorting cells. These activities, we have determined, are being facilitated by flow cytometry. At Bio-Rad, we have incorporated flow cytometry into our CRISPR workflows. Be- sides monitoring transfection and enriching cell populations, we have been confirming edits and conducting downstream analyses. By pairing multiple tools and techniques, we have increased workflow efficiency. At the end of the day, scientists using CRISPR want to know that they have successfully edited their target cells before moving to down- stream assays. GEN: Now that flow cytometry is being adapted for immunology, will new applications emerge to advance immunotherapy? Dr. Guenther: As the immunotherapy field continues to ex- pand, flow cytometry will play an important role in the development of varied immuno- therapies. I see the method mostly involved in the quality control of the cell validation process during and after treatment, and in the monitoring of the therapeutic cells in the patient over time. Many cellular subsets and functional markers of immune cells can be rapidly and efficiently analyzed by flow cytometry with- out the need of additional assays, so flow cytometry can also be applied to predict off- target effects. Finally, an exciting emerging area is the integration of flow cytometry into multi-omics for the application of precision medicine in immuno-oncology. Ms. Wright: Traditional immunology was grounded in the principles of immune function, both in- nate and acquired. Immunotherapy is no different, and we'll need to return to these principles to understand what a tumor does to evade the immune response and which aspects of the innate and adaptive immune systems can be harnessed to enable tumor elimination or avoidance altogether. Some fascinating science is taking place in the immunotherapy field, particularly a focus on understanding the tumor micro- environment. This is putting a spotlight on flow cytometry as well as new technologies such as acoustically focused flow cytometry. These techniques are important in this re- search area because they can handle samples derived from tissues without complications such as clogging, and they enable efficient analysis of rare cell populations. Ms. Talaga: Chronic viral infections and cancers have always tried to evade the immune system. Today, we are recognizing that the immune system cannot be studied as though it ex- isted in isolation. Instead, its relationship to the microbiome must be recognized, as well as its (potential) vulnerability to combined therapies. We have been learning to reach across scientific disciplines and exchange both knowledge and techniques. This combination of interdisciplinary work and knowledge sharing has led, in turn, to what we call the "emerging era of immunotherapies." Scientists who incor- porate flow cytometry into greater work- flows—such as workflows encompassing PCR detection—are finding greater success. For example, PCR detection may be fol- lowed by electroporation and flow cytom- etry, allowing us to acquire new knowledge about, say, stem cells, and apply it into clini- cal work. " We can now analyze dozens of 'markers,' both proteins and nucleic acids, with single- cell resolution and simultaneously derive valuable population statistics." — Dara Grantham Wright, Thermo Fisher Scientific HTsupport © 2018 Promega Corporation. 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