InquiryCRISPR gene editing technology has been used as a tool in the fields of biological discovery and innovative drug discovery for screening. For example, while using the CRISPR technology for gene screening (pooled CRISPR screens), cells can be edited simultaneously by CRISPR targeting to hundreds of different genes and screening for gRNAs in edited cells. Meanwhile, gRNAs can help determine which of the genes are critical for biological mechanisms of action. A new technology, CROP-seq (CRISPR droplet sequencing), creatively combines the CRISPR screening technology with the single-cell RNA-seq technology to analyze single-cell transcription profiling of target genes that are edited by gRNA. CROP-seq uses high-throughput sequencing to output more complex regulatory information, bringing unprecedented improvement to the research of gene regulatory mechanisms and drug target screening.
Our CROP-Seq Platform
CD Genomics has launched a next-generation sequencing platform – CROP-Seq, for single-cell, with an aim to help researchers analyze and validate genome editing with high throughput.
CROP-seq screening[1]
Applications
1. High-throughput screening and analysis of gene regulatory functions
2. Study of complex signaling pathways, i.e., physiological and pathological processes
3. Study of complex genetic and epigenetic regulation, i.e., cellular processes
4. High-throughput drug target screening
Technical Advantages
1. Adopted the CRISPR hybrid library screening (pooled CRISPR screens) method, large-scale and efficient screening of genes of action can be achieved.
2. Using the unique CRISPR vector, the single-cell transcriptome can be directly detected via gRNA to greatly simplifies screening steps.
3. Utilizing single-cell transcriptome sequencing to output more data and information, providing complex gene regulation maps.
| Basic Analysis | Advanced Analysis |
|---|---|
| 1. Raw data quality control | 1. Differential gene analysis of gRNA expression between groups |
| 2. Cell number identification | a) Differential gene screening between groups |
| 3. Genome alignment | b) Differential gene screening among multiple groups |
| 4. Counts calculation | 2. WGCNA analysis of gRNA expression between groups |
| 5. Cell filtration | a) Functional enrichment analysis |
| 6. Single-cell expression screening | b) Key genes screening |
| 7. gRNA distribution | c) Gene function prediction |
| 8. PCA | 3. Pathway analysis of gRNA cell expression between groups |
| a) An overview of classical pathways, upstream regulatory molecules, disease, biological functions, etc. | |
| b) Network gene interactions: finding activating and repressing genes in pathways Comparative analysis of biological functions of enrichment pathways | |
| d) Prediction of upstream regulatory genes and downstream targets |
Multiple CRISPR/Cas9 CROP-Seq Libraries Available
RNA binding protein CRISPR/Cas CROP-Seq library
Epigenetic regulator CRISPR/Cas CROP-Seq library
Transcription factor CRISPR/Cas CROP-Seq library
Custom CRISPR/Cas CROP-Seq library
Sample Requirements
Specie: human only
Type: frozen cells, live cells
References
- CROP-Seq: a single-cell CRISPRi platform for characterizing candidate genes relevant to metabolic disorders in human adipocytes. bioRxiv 2022
- Datlinger P, Rendeiro AF, Schmidl C, et al. Pooled CRISPR screening with single-cell transcriptome readout. Nat Methods 2017 03;14(3).
