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
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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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New Tool Enables Researchers to Rapidly Manipulate Protein Levels in Mammalian Cells 

New Tool Enables Researchers to Rapidly Manipulate Protein Levels in Mammalian Cells  | Genetic Engineering Publications - GEG Tech top picks | Scoop.it

http://www.nature.com/ncomms/2016/160527/ncomms11742/full/ncomms11742.html

BigField GEG Tech's insight:

A research team headed by investigators at the Icahn School of Medicine at Mount Sinai (ISMMS) has developed a tool that allows scientists to quickly manipulate levels of two proteins in the same cell. They say the method, dubbed a “dual molecular tuner,” offers an easy way to perform in-depth analyses in mammalian cells in general, and stem cells in particular.

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New tools to manipulate biology 

New tools to manipulate biology  | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
BigField GEG Tech's insight:

Chemistry has provided many key tools and techniques to the biological community in the last twenty years. We can now make proteins that Mother Nature never thought of, image unique parts of live cells and even see cells in live animals. In ACS Central Science, three independent research groups from the University of Geneva (UNIGE) and one from the University of Basel (UNIBAS) take these accomplishments a step further, reporting advances in both how proteins are made and how you can see their expression patterns in live animals.

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