Single-Cell CRISPR Screening Service: Perturb-Seq, CRISP-Seq, CROP-Seq etc.
With Cellecta’s Single-Cell CRISPR Screening Service (aka Perturb-Seq, CRISP-Seq, CROP-Seq, etc.) you can study the effect of CRISPR-based perturbations on mRNA expression in many thousands of cells using single-cell RNA sequencing (scRNA-Seq) on the 10X Chromium system. This method can provide important insights into the transcriptional response resulting from genetic mutations and other perturbations that affect gene function.
Investigate transcriptional changes associated with 100+ genes in a single experiment
Combine pooled CRISPR knockout (KO), CRISPR inhibition (CRISPRi) or CRISPR activation (CRISPRa) screening with single-cell expression analysis
Identify genes affecting gene and pathway activation that are missed with conventional arrayed screens
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General Information
Cellecta offers its Single-Cell CRISPR Screening Service on the 10X Chromium Controller Platform, which allows for screening of multiple genes in a single experiment and enables the identification of transcriptional profiles linked to specific gene perturbations.
We carry out all the services required to run most screens onsite, including construction and packaging of custom pooled sgRNA libraries, single-cell RNA-Seq on our in-house 10X Chromium and Illumina sequencing instruments, alignment of reads to identify hits, and post-screening analysis.
Single-Cell CRISPR Screening workflow
Step 1: Clone sgRNA library
Step 2: Cells transduced with pooled sgRNA libraries are loaded in the 10X reaction
Step 3: Beads are co-encapsulated with unique barcodes and cells are dispersed in discrete droplets
Step 4: The sgRNA and mRNAs labeled with bead barcodes are used to construct libraries and running scRNA-Seq, single-cell transcriptional profiles are generated
Step 5: Changes in gene expression associated with specific sgRNAs are linked to gene-specific knockouts
In addition to the full-service screening experiment, you may be interested in:
Dixit A, Parnas O et al. (2016) Perturb-Seq: Dissecting Molecular Circuits with Scalable Single-Cell RNA Profiling of Pooled Genetic Screens.Cell 167( 7): 1853–1866. [abstract]
Adamson B, Norman TM et al. (2016) A Multiplexed Single-Cell CRISPR Screening Platform Enables Systematic Dissection of the Unfolded Protein Response.Cell 167( 7):1867–1882. [abstract]