Genetic Engineering Publications - GEG Tech top picks
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CRISPR-FOCUS: A web server for designing focused CRISPR screening experiments

CRISPR-FOCUS: A web server for designing focused CRISPR screening experiments | Genetic Engineering Publications - GEG Tech top picks | Scoop.it

CRISPR-FOCUS is freely available at http://cistrome.org/crispr-focus/

BigField GEG Tech's insight:

The recently developed CRISPR screen technology, based on the CRISPR/Cas9 genome editing system, enables genome-wide interrogation of gene functions in an efficient and cost-effective manner. Although many computational algorithms and web servers have been developed to design single-guide RNAs (sgRNAs) with high specificity and efficiency, algorithms specifically designed for conducting CRISPR screens are still lacking. Here the scientists present CRISPR-FOCUS, a web-based platform to search and prioritize sgRNAs for CRISPR screen experiments. With official gene symbols or RefSeq IDs as the only mandatory input, CRISPR-FOCUS filters and prioritizes sgRNAs based on multiple criteria, including efficiency, specificity, sequence conservation, isoform structure, as well as genomic variations including Single Nucleotide Polymorphisms and cancer somatic mutations. CRISPR-FOCUS also provides pre-defined positive and negative control sgRNAs, as well as other necessary sequences in the construct (e.g., U6 promoters to drive sgRNA transcription and RNA scaffolds of the CRISPR/Cas9). These features allow users to synthesize oligonucleotides directly based on the output of CRISPR-FOCUS. Overall, CRISPR-FOCUS provides a rational and high-throughput approach for sgRNA library design that enables users to efficiently conduct a focused screen experiment targeting up to thousands of genes.

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Evaluation of off-target and on-target scoring algorithms and integration into the guide RNA selection tool CRISPOR

Evaluation of off-target and on-target scoring algorithms and integration into the guide RNA selection tool CRISPOR | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
BigField GEG Tech's insight:

Here the scientists conduct the first independent evaluation of CRISPR/Cas9 predictions. To this end, they collect data from eight SpCas9 off-target studies and compare them with the sites predicted by popular algorithms. They find that the optimal on-target efficiency prediction model strongly depends on whether the guide RNA is expressed from a U6 promoter or transcribed in vitro. They further demonstrate that the best predictions can significantly reduce the time spent on guide screening.

 

To make these guidelines easily accessible to anyone planning a CRISPR genome editing experiment, they built a new website (http://crispor.org) that predicts off-targets and helps select and clone efficient guide sequences for more than 120 genomes using different Cas9 proteins and the eight efficiency scoring systems evaluated here.

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