Yorba Linda, CA (PRWEB) September 02, 2015
The CRISPR-Cas9 system introduces double-strand DNA breaks at a specific locus in the genome by using a complex of the Cas9 nuclease with either a chimeric single guide RNA (sgRNA) or two short RNAs (a CRISPR RNA (crRNA) and a trans-activating RNA (tracrRNA)). The ability of any given guide RNA (sgRNA or crRNA) to create a break in the target DNA that causes functional protein disruption can vary based on the guide RNA (gRNA) sequence and position in the targeted gene. Likewise, the overall specificity of RNA-directed DNA cleavage events is not yet completely understood and can hamper its wider application.
Dharmacon, part of GE Healthcare, presents a new educational webinar, “Improve CRISPR-Cas9 experiments with rationally designed guide RNAs,” which will discuss a new algorithm to score gRNA sequences based on how likely they are to produce functional knockout of targeted genes. The speaker is Alex Amiet, Senior Product Manager for Dharmacon Gene Editing, GE Healthcare.
While gRNAs targeting one or a few genes can often be chosen ad hoc, performing high throughput genome-scale loss-of-function screens requires gRNAs that have consistently high functional knockout efficiencies. To understand the parameters affecting CRISPR-Cas9 gene editing efficiency, GE systematically evaluated over 1100 synthetic crRNAs in a reporter assay and identified characteristics important for functional gene disruption. Using this data, they developed an algorithm to score gRNA sequences based on how likely they are to produce functional knockout of targeted genes. The algorithm was tested by designing synthetic crRNAs to genes unrelated to the original assay and examined their ability to knock out gene function using additional phenotypic assays. Results demonstrate that high-scoring crRNAs have increased functionality. Further, GE developed an optimized alignment program to perform complete specificity analysis of crRNAs, including detection of gapped alignments. Recent work has demonstrated gene editing by crRNAs containing bulges of up to 4 nucleotides, but a number of existing design tools are unable to detect putative off-targets based on gapped alignments.
Attendees who join will learn about:
- The importance of a thorough and accurate guide RNA specificity alignment
- How rational design of guide RNAs can improve CRISPR functionality
- The impact of genome-wide, algorithm-designed guide RNAs on gene editing efficacy and time to results
A live question-and-answer session will follow the presentation.
Alex Amiet is a Global Senior Product Manager at Dharmacon, part of GE Healthcare. She is responsible for developing and expanding the Edit-R™ Genome Engineering portfolio, working closely with a renowned team of R&D scientists to bring to market innovative CRISPR-Cas9 tools and services. In her ten-year tenure at Dharmacon, she has built and led the development of the novel portfolio of microRNA research tools from the ground-up and managed nearly the entirety of the Dharmacon product lines, including custom RNA and siRNA synthesis, shRNA, gene expression and genome-scale RNAi screening services for drug development. Alex studied Molecular Biology at the University of Houston and has developed new Life Science technology platforms for Thermo Fisher Scientific, Sigma-Aldrich and Degussa GmbH for over 16 years.
The free webinar, hosted by LabRoots, will be presented on September 29, 2015, at 8 am PT.
For full details and free registration, click here.
Leveraging expertise in bioinformatics, RNA biology, and synthesis chemistry, the company has developed a complete line of products for the RNAi researcher. Further breakthroughs arose in chemical modifications for siRNA specificity, and novel molecules for microRNA modulation. In 2014, Dharmacon became part of GE Healthcare Life Sciences. For more information visit gelifesciences.com/dharmacon
LabRoots is the leading professional networking website designed to connect all science verticals. Founded in March 2008, LabRoots’ vision was to connect the scientific world leveraging a myriad of unique features and tools, discovering meaningful collaborations across geographic boundaries and fields of expertise. LabRoots is the owner and producer of BioConference Live – which has grown into the world’s largest series of virtual events within the Life Sciences and Clinical Diagnostics community.