Genetic Engineering & Biotechnology News

DEC 2017

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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12 | DECEMBER 2017 | ➜ DeeAnn Visk, Ph.D. The CRISPR/Cas9 system, we are told, is so very easy to implement— and it is, at least initially. But CRISPR/ Cas9 may soon run into difficulties. For example, a CRISPR/Cas9-based process may need to be scaled up or streamlined, reoriented toward more sophisticated tasks, or refined to deliv- er consistent, verifiable results. When that happens, a CRISPR/Cas9 process may betray its improvisational origins or its jerry-rigged nature. Fortunately, CRISPR/Cas9 is be- coming more systematized through the introduction of off-the-shelf tools: oligo libraries, assay kits, selectable markers, and more. These tools, in the hands of creative investigators, enable increasingly powerful genome-engi- neering platforms—more precise, more efficient, and more capable of deliver- ing modifications that show superior fit and finish, as may be determined with —you guessed it—commercially available assays. A prominent provider of CRISPR/ Cas9 tools is GE Healthcare Dharma- con. The company's products support applications from CRISPR/Cas9 gene editing to functional genomics. "The entire human genome is available as synthetic CRISPR RNAs (crRNAs) in a 96-well or 384-well plate format, which can easily be used in arrayed phenotypic screens," states Annaleen Vermeulen, Ph.D., a senior scientist at GE Healthcare Dharmacon. "We design and synthesize crRNAs to target every gene in the genome, and each crRNA is provided in its own individual well." The approach permits a pheno- typic readout of cell response using an automated microscope. Earlier this year, an example of this approach was described in the Journal of Biotechnol- ogy. Essentially, 169 genes pertinent to cell-cycle regulation were targeted for disruption with CRISPR/Cas9 reagents delivered by lipid transfection. Seven different parameters of the cells were measured in a high-content assay. "One advantage of the system of- fered by Dharmacon is that we have designed at least four different guide RNAs per gene," asserts Dr. Vermeu- len. "When these different guide RNAs are utilized, the gene to be disrupted is targeted at four different loci. Thus, if the same effect is seen using all four of the guide RNAs for the same gene, then the results are more likely to be biologically relevant and not merely an artifact of the CRISPR/Cas9 editing or an off-target effect." Genome Engineering: CRISPR Proving More User-Friendly B E S T O F C R I S P R 2 017 Scaling CRISPR Technology to Produce Off-the-Shelf Libraries Deriving an edited monoclonal cell line: Typical workflow with CRISPR editing. Advanced Analytical (AATI)

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