DATE: July 25, 2017
TIME: 9:00am PT, Noon ET
Thermo Fisher Scientific is proud to present the SyncD3 webinar series. As a thought-leader in science our first commitment is to you the scientist and partnering to drive the conversation on current practices, standards and the development of innovation.
Join us for our continuation of the SyncD3 webinar series and virtual event, where we will discuss cutting-edge tools and topics in the field(s) of Drug Discovery & Development such as:
We will discuss where these areas intersect and impact drug discovery and development and the future of the industry during a live panel discussion with our presenters and you the audience. Learn from real field scientists and researchers who are working in the ADME/Tox and Drug Discovery fields and bring your questions for the open forum Q&A session after our presentations.
If you cannot make the live presentation you are always capable of accessing the recorded information, presentations and Q&A any time after the event by registering here.
Unraveling biology and identifying targets with functional genomics approaches supported by LentiArray CRISPR library screens
Identifying and validating targets that underlie disease mechanisms and can be addressed to provide efficacious therapies remains a significant challenge in the drug discovery and development process. Mechanisms of RNAi have provided the use of siRNA and shRNA to knock-down RNA and suppress gene function. However, depending on the nature of the targets, cells, biology and end-point assays, these approaches may suffer variously from their transient nature, design complexity, incomplete knock-down or off-target effects. The use of CRISPR (clustered regularly interspaced short palindromic repeat)-associated Cas9 nuclease and guide RNA (gRNA) provides a strong alternative that can produce transient or long-lasting impact, straightforward design, knock-out of genes and increased specificity. A number of laboratories have already published reports demonstrating how pools of gRNA can be delivered to cells and “hits” can be established through enrichment or depletion of cells following a “survival” assay and identified by sequencing the introduced gRNAs in the remaining cell population. Here we demonstrate a knock-out screening approach that utilizes the Invitrogen™ LentiArray™ CRISPR library to interrogate the impact of individual gene knock-outs on the NFκB pathway as measured by a functional cell-based assay. We describe the library design concepts, the assay development, initial screening results and validation of specific identified hits. We elucidate the key factors in developing a robust assay including both transduction and assay optimization to achieve the highest levels of transduction efficiency and assay window and provide data from initial screens using the Invitrogen™ LentiArray™ CRISPR kinome library. We expect these approaches to be scaleable to the entire human genome and portable to multiple cell types and end-point assays including both high-throughput plate-based assays and high-content imaging based assays.