RNAi is a common approach to genome screening today, however it has challenges with incomplete functional knockdown, low target efficacy, false negatives, increased toxicity and cell type limitations due to viral delivery. This results in increased costs and downstream validation as well as important missed targets.
While pooled lentiviral CRISPR screening addresses some RNAi challenges, it’s not ideal as it introduces large numbers of guides into cells, and limits assays to few screening phenotypes such as cell growth and death. Pooled lentiviral screening also poses logistical challenges working with live virus, guide representation bias, variable editing efficiencies, and complicated data deconvolution through next-generation sequencing (NGS).
Synthego libraries supercede RNAi and pooled CRISPR screening by combining the advantages of an arrayed format, high-quality target designs, modified synthetic sgRNA, and whole genome coverage. The arrayed sgRNA libraries arrive ready-to-transfect in any human cell type and enable straightforward testing, assaying and analysis of complex phenotypes beyond cell viability, allowing for identification of targets involved in complex diseases such as neurodegeneration and cardiovascular disease.
By using modified sgRNA during the screening phase, researchers’ results translate directly to the next phases of development without any need to change guide formats. Furthermore, arrayed screening with modified synthetic sgRNA eliminates aforementioned logistical challenges with an automation-friendly, virus-free, transfection-ready sgRNA library that provides consistent editing efficiencies without complicated NGS.
“Arrayed functional screening with high quality modified synthetic sgRNA libraries simplifies development and enables scientists to rapidly screen in an unbiased manner for relevant phenotypes in any cell type, including human stem cells and primary cells,” said Abhi Saharia, PhD, director of product management. “This paves the way for the advancement of drug discovery and medical applications through faster and more accurate identification of new drug targets and better understanding of the gene function.”
The Synthego whole human genome library is available in gene families including the druggable genome, transcription factors, G-protein coupled receptors, kinases, and immunology and immuno-oncology targets, among others. This important feature enables focused CRISPR screens, allowing hypothesis-driven gene targeting, and running of simultaneous screens without overlap.
Designed with the Synthego CRISPR Design Tool, modified synthetic sgRNAs in the library ensure a comprehensive CRISPR screen with the greatest gene knockout efficiency. These guides target every protein-coding gene in the human genome and achieve up to 90 percent editing efficiency with high consistency across all human cell types, enabling unbiased whole genome screening even in human primary and stem cells.
For a full list of the gene families visit: www.synthego.com/libraries
For more information on Synthego visit: www.synthego.com.