CLUE

CLUE generates multiplexed custom single-guide RNA (sgRNA) libraries by combining bioinformatics design and a barcoded oligonucleotide pooling strategy to enable tailored CRISPR/Cas9 perturbation experiments.

Key Features:

• Custom Library Design: Designs sgRNA libraries from user-provided gene or sequence lists and encodes them into a single synthetic oligonucleotide pool.
• Multiplexing Capability: Employs a barcoding strategy that assigns unique primer binding sites to distinct libraries within one oligonucleotide pool, enabling simultaneous amplification of multiple sgRNA libraries.
• Uniformity and Specificity: Ensures uniform distribution of sgRNAs across multiplexed libraries while minimizing cross-contamination between libraries.
• Resource Efficiency: Enables parallel production of numerous custom sgRNA libraries from a single oligonucleotide pool to reduce reagent and synthesis resources.

Scientific Applications:

• CRISPR/Cas9 Perturbation Screens: Generates tailored sgRNA sets for systematic genome perturbation experiments using CRISPR/Cas9.
• Complex and In Vivo Studies: Supports experiments requiring specific gene sets under complex biological conditions, including in vivo systems.

Methodology:

Performs in silico design of sgRNAs from input gene or sequence lists and assigns barcodes as unique primer binding sites to multiplex libraries within a single synthetic oligonucleotide pool.