Combinatorial CRISPR technologies have emerged as a transformative approach to systematically probe genetic interactions and dependencies of redundant gene pairs. However, the performance of different functional genomic tools for multiplexing sgRNAs vary widely. Here, we generate and benchmark ten distinct pooled combinatorial CRISPR libraries targeting paralog pairs to optimize digenic knockout screens. Libraries composed of dual Streptococcus pyogenes Cas9 (spCas9), orthogonal spCas9 and Staphylococcus aureus (saCas9), and enhanced Cas12a from Acidaminococcus were evaluated. We demonstrate a combination of alternative tracrRNA sequences from spCas9 consistently show superior effect size and positional balance between the sgRNAs as a robust combinatorial approach to profile genetic interactions of multiple genes.
Information superiority is a strategic goal of the US Air Force. To achieve this goal the Air Force aims to produce a battlespace infosphere that will provide an unprecedented degree of connectivity and availability of raw data and value-added information for warfighter use. The
AbstractInformation superiority is a strategic goal of the US Air Force. To achieve this goal the Air Force aims to produce a battlespace infosphere that will provide an unprecedented degree of connectivity and availability of raw data and value-added information for warfighter use. The essential challenge of the infosphere is to be able to provide the right information, at the right time, in the right form to enable warfighters to take effective, coordinated action. Although the infosphereÕs core web and agent technologies are clearly able to provide a heterogeneous infosphere, improved interface technologies are also needed to address problems of information overload and how to provide support to specific end-users without the support tools themselves becoming an impediment to task performance. We have developed a prototype WorkCentered Support System software client as a means to address these interface issues. The WCSS approach achieves effective support in a software agent environment by blending direct manipulation, work field organization, and decision, collaborative, and product development aiding in a manner that is tailored to both formal and informal characteristics of user work. In this paper we describe the philosophy behind and characteristics of the WCSS technology. We illustrate the technology with a discussion of an interactive WCSS prototype designed to improve support to military airlift mission planners at the headquarter level.
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