Single-Cell CRISPR Screens. Perturb-Seq, CROP-Seq and more. Simplified.

Pooled CRISPR screening with lentiviral sgRNA libraries has proven to be an effective approach to identify genes functionally required to generate particular phenotypes, and RNA-Seq is an effective method to find the underlying changes in gene expression producing those phenotypes. These two single-cell CRISPR screening methods, often called Perturb-Seq or CROP-Seq assays, may be combined to assess and characterize the distinct expression profiles produced by a genetic disruption in a pooled CRISPR knockout screen.
We are pleased to announce that Cellecta now offers a Single-Cell CRISPR Screening Service. We can run your Perturb-Seq, CROP-Seq or other single-cell screening method of choice with our in-house 10X Chromium and Illumina NGS instruments.

How it works

Single-Cell CRISPR Screening workflow

Step 1: Clone sgRNA library
Step 2: Cells transduced with pooled sgRNA libraries are loaded in the 10X reaction (10X Chromium platform)
Step 3: Beads are co-encapsulated with unique barcodes and cells are dispersed in discrete droplets
Step 4: The sgRNA and mRNAs labelled 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

More about Cellecta Single-Cell CRISPR Screening Service

Are you looking to do a pilot study or run your own single-cell CRISPR screen?
Cellecta offers products to support your do-it-yourself CROP-Seq or Perturb-Seq projects

CloneTracker^TM XP-CRISPR Human and Mouse Anti-Cancer CRISPR Barcode Libraries
Custom CloneTracker XP CRISP-Seq Libraries
CloneTracker^TM Barcode Libraries and Kits

1. CloneTracker^TM XP-CRISPR Human or Mouse Anti-Cancer CRISPR Barcode Libraries

Ready-to-use libraries targeting 27 human anti-cancer genes or 27 corresponding mouse homologs. There are 20 sgRNAs to each gene designed to target functional domains. This is ideal if you are running a Single-cell Screen yourself or want to try to do so with a smaller library.

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2. Custom CloneTracker XP CRISP-Seq Libraries

Cellecta provides custom-designed CRISPR sgRNA Pooled Libraries targeting virtually any set of genes and incorporating guide sequences and vector features for a range of screening applications.
We offer two library formats compatible with single-cell analysis:

The sgRNA-encoding oligos are cloned into one of our pScribe vectors so the guide sequences are expressed and incorporated into the cDNA where they are detectable during single-cell expression profiling.
Alternatively, sgRNA that includes a 10X feature barcode capture sequences in the tracr domain of the sgRNA can be captured during single-cell analysis profiling on a 10X instrument.

Simply let us know the CRISPR libraries your study requires, and let our experts produce them for you.

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3. CloneTracker^TM Cell Barcoding Libraries and Kits

Label whole cell populations with stable, genomically integrated, single, sequenceable barcodes for cell tracking and quantification of descendants in mixed downstream cultures or tissue. Library constructs integrate into the genomic DNA after transduction and can be detected by genomic DNA amplification and RNA sequencing analysis.

Uniquely label millions of cells with our ready-to-use or custom libraries
Identify gene activation changes in response to knockouts on a cell-to-cell basis
Assess responses of each of the clonal progeny from the initial barcoded population
Determine how experimental conditions, differentiation, or developmental processes affect distinct groups of cells
Off-the-shelf, ready-to-use libraries are available immediately.

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Additionally, Cellecta can also provide custom lentiviral barcode libraries that enable you to generate populations of millions of cells where each has its own uniquely identifiable barcode sequence according to what your study requires.
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Also available are Individual Barcode Panels to label all the cells in a defined population with one specific barcode so they and their progeny can be identified when mixed and co-cultured with other cell populations.

Related Publications:

Adamson, B., et al., A Multiplexed Single-Cell CRISPR Screening Platform Enables Systematic Dissection of the Unfolded Protein Response. Cell 167: 1867–1882.e21 (2016)

Datlinger, P., et al., Pooled CRISPR Screening with Single-Cell Transcriptome Read-Out . Nat Methods 14: 297–301 (2017)

Dixit, A., et al., Perturb-Seq: Dissecting Molecular Circuits with Scalable Single-Cell RNA Profiling of Pooled Genetic Screens. Cell 167: 1853–1866 (2016)

To learn more about other CRISPR screening offerings as well all of Cellecta products and services, visit www.cellecta.com or email us at info@cellecta.com

Single-Cell CRISPR Screening.

Pooled Genetic Screening and RNA Expression Profiling at the Single-Cell Level.