CRISPR-FOCUS: A web server for designing focused CRISPR screening experiments

Area of Science:

• Genomics
• Molecular Biology
• Bioinformatics

Background:

• CRISPR-Cas9 technology enables efficient genome-wide gene function studies.
• Existing tools for designing single-guide RNAs (sgRNAs) lack features specific to CRISPR screens.

Purpose of the Study:

• To develop a web-based platform, CRISPR-FOCUS, for searching and prioritizing sgRNAs for CRISPR screen experiments.
• To provide a rational and high-throughput approach for sgRNA library design.

Main Methods:

• CRISPR-FOCUS filters and prioritizes sgRNAs based on efficiency, specificity, sequence conservation, isoform structure, and genomic variations (SNPs, somatic mutations).
• The platform accepts official gene symbols or RefSeq IDs as input.
• It also provides control sgRNAs and necessary construct sequences (promoters, scaffolds).

Main Results:

• CRISPR-FOCUS offers a comprehensive solution for designing sgRNA libraries for focused CRISPR screens.
• The platform facilitates direct oligonucleotide synthesis based on its output.
• It enables efficient screening of thousands of genes.

Conclusions:

• CRISPR-FOCUS provides a valuable resource for researchers conducting CRISPR screens.
• The platform streamlines the design process, enhancing the efficiency and accuracy of gene function studies.
• It supports high-throughput screening by rational sgRNA selection.