Your trusted partner in CRISPR-based functional genomic screening
of human cells
Choose the CRISPR-Ready ioCell type for your screen
We currently have CRISPRko-Ready ioGlutamatergic Neurons, CRISPRko-Ready ioMicroglia, CRISPRa-Ready ioGlutamatergic Neurons and CRISPRi-Ready ioGlutamatergic Neurons, which can be used in our screening platform. If you are interested in performing a screen in a different ioWild Type Cell or ioDisease Model Cell, speak with us about generating the right model system for your screen.
Nominate your genes of interest
Study the effect of knocking out tens to thousands of genes in one screen. Whether you want to study a particular pathway or perform a genome-wide screen, our expert team of scientists and bioinformaticians are here to design a custom sgRNA library based on your needs.
Choose your readout
With a range of readouts including high-throughput viability assays, fluorescence-based phenotypic screens, next-generation sequencing (NGS) to identify gene knockout effects and single-cell RNA sequencing, our platform enables you to discover key genetic drivers of biological processes. We provide the tools and expertise to deliver high-resolution, data-rich insights tailored to your research needs.
Receive your data
Need more support? We offer full data analysis support for your CRISPR screening results.
Functional readouts
from CRISPR screens
Functional readouts
from CRISPR screens
A pooled single cell CRISPR knockout screen uncovers modulators of microglia activation.
To conduct a pooled single cell CRISPR knockout screen (scCRISPR screen) with a targeted sequencing readout, we first identified a transcriptomic activation signature of 258 differentially expressed genes by comparing CRISPR-Ready ioMicroglia treated with and without LPS at day 10. Separately, we selected 110 candidate genes for the pooled scCRISPR screen based on their known roles in neurodegeneration and neuroinflammation. Guide RNAs were delivered via lentiviral transduction on day 10, aiming for a single integration per cell. The cells were cultured and then treated with +/- LPS for 24 hours before single cell processing on day 15. Cosine similarity analysis compared knockouts in LPS-treated CRISPR-Ready ioMicroglia to both resting and activated states. The analysis identified 17 gene knockouts that altered responses to LPS stimulation. The heatmap shows Log2FC profiles for gene knockouts that had a cosine similarity above 0.3 (arbitrarily chosen threshold) compared to cells with non-targeting guides in the unstimulated condition. Knockouts are sorted based on their cosine similarity to the non-LPS condition. CD14, MAP3K7, TIRAP, IKBKG, TRAF6, IKBKB, LY96, TICAM1, RELA, and TLR4 are genes known to be involved in LPS activation mediated via the TLR4 signalling pathway.
A pooled knockout screen of neurodegenerative disease-relevant genes in CRISPR-Ready ioGlutamatergic Neurons shows clustering of aaRS genes in UMAPs
For a pooled knockout screen in CRISPR-Ready ioGlutamatergic Neurons, 100 known genes involved in neurodegenerative diseases were selected. Lentiviral transduction of the gRNAs was carried out on day 3, and single-cell gene expression analysis was performed on day 12. Single cells were clustered on uniform manifold approximations and projections (UMAPs) based on their shared nearest neighbour’s gene expression. Clustering of aminoacyl-tRNA synthetase (aaRSs) knockouts including AARS1, HARS1, CARS1, and GARS1 was observed. In contrast, cells transduced with non-targeting control sgRNAs were evenly distributed among clusters. Pathway analysis showed gRNAs targeting aaRSs activated the unfolded protein response (UPR), the mechanism by which cells control endoplasmic reticulum protein homeostasis. In many neurodegenerative diseases, signs of UPR activation have been reported. The most common aaRS-associated monogenic disorder is the incurable neurodegenerative disease Charcot–Marie–Tooth neuropathy (CMT).
Custom CRISPR screens and CRISPR-Ready ioCell development
If we don't currently have the cells your target identification and validation workflows need, our team of experts can develop new CRISPR-Ready ioCells in any ioWild Type Cell background. Additionally, if you are looking to outsource the development of high throughput CRISPR screens in CRISPR-Ready ioCells, we can design, build and run these for you.
