Whole Genome Synthetic sgRNA Libraries

CRISPR Screening for Comprehensive Target Identification in Human Cell Lines

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Genome-wide loss-of-function screens have been incredibly valuable in identifying novel drug targets and in uncovering the functional complexity of the human genome. This approach had largely been limited to RNAi (shRNA and siRNA) or pooled CRISPR screening. Although valuable, RNAi presents several challenges including incomplete functional knockdown and high false negatives.

Pooled CRISPR screening addresses some of these issues, but presents other logistical challenges such as guide representation bias, limited testable phenotypes and complicated data deconvolution through next-gen sequencing.

Summary

Synthego’s modified synthetic single guide RNA (sgRNA) libraries for arrayed whole human genome CRISPR screening solves these problems with an automation-friendly, virus-free sgRNAs that provides consistently high editing efficiencies without complicated NGS. The sgRNAs are designed with the Synthego Design Tool to maximize gene knockout by targeting an early exon that is common to all known mRNA transcripts, and minimize off-target effects by selecting guides that have minimal off-target sites in the human genome. The arrayed sgRNA libraries arrive ready-to-transfect in any human cell type, including primary and stem cells, and enable easy testing of multiple complex phenotypes beyond cell viability.

The Synthego whole human genome library is available in gene families including the druggable genome, transcription factors, G-protein coupled receptors, kinases and immunology and immuno-oncology targets, among others.

Screen better, discover quicker and elevate your screening with the best-in-class genome-wide screening library!

Benefits

Comprehensive
Average of 3 sgRNAs for all the protein-coding genes in human genome

Well-designed
Best-in-class guide design through the Synthego Design Tool

Effective
Modified synthetic sgRNA for high efficiency knockouts

Flexible
Enables any phenotypic assay, e.g., imaging, fluorescence or luminescence

Easy-to-Use
Automation-friendly 384-well plates, ready-to-transfect sgRNA

Powerful
Accurate hit identification without NGS

Library Specifications

– Human only
– Library constructed in 384-well barcoded plates
– Each well contains 3 multiplexed synthetic modified sgRNA targeting a single gene
– Each synthetic guide is verified by mass spectrometry
– Negative controls included on each plate
– Space included for biological and technical positive controls per plate

Available Libraries

Whole Genome
Apoptosis Pathway
B-Cell Activation
Cell Adhesion Genes
Cell Cycle Regulators
Current Drug Targets
Cytokine and Chemokine
Cytoskeleton Genes
Deubiquitinating Enzymes
DNA Repair Pathway
Druggable Genome
Epigenetic Regulators
Extracellular Matrix Genes
G-Protein Coupled Receptors
Helicase
Immunology/Immuno-Oncology
Ion Channel
JAK-STAT Pathway
Kinases, Complete
Nuclear Hormone Receptors
p53 Pathway
Phosphatases
Serine Proteases
T-Cell Activation
Transcription Factors
Tumor Suppressors
Tyrosine Kinases
Ubiquitin Ligases (E1, E2, E3)

High Editing Efficiencies in a Kinase Library

HEK293 Cells

HEK293

Jurkat Cells

Jurkat

Four guide RNAs were designed per gene to knockout 9 kinases across the human genome. These were constructed as chemically modified synthetic sgRNA and electroporated into either HEK293 cells or Jurkat cells. The DNA from the resulting pool of cells was analyzed by TIDE analysis, demonstrating a high editing efficiency across the board with at least one guide per gene for every gene tested, giving greater than 50% editing efficiency (red line denotes 50% editing efficiency). This demonstrates the high efficiency with which guides designed and constructed by Synthego perform within a library.

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