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single cell crispr screening

Single-Cell CRISPR Screening CapitalBio

Traditional CRISPR screening is to first construct a mixed sgRNA library, which can contain hundreds to thousands of sgRNAs, and then transfect the library into cells that can stably express Cas9 protein to achieve large-scale gene knockout. Cells are subjected to phenotype screening and bulk sequencing to obtain information on sgRNA changes during the culture process, so as to know the impact of the knocked-out gene on the cells. This conventional method can only screen macroscopic phenotypes of cells, so it is more suitable for answering questions directly related to cell survival or macroscopic phenotypes, but not suitable for research on molecular phenotype changes. Based on this, 10x Genomics has developed a single-cell CRISPR screening platform, which uses high-throughput single-cell sequencing to obtain the gene expression profile and sgRNA expression of a single cell from the cells screened by CRISPR. The results of the CRISPR screen were tested at the level of phenotype.


Method Applications Analysis Display

Figure1.jpg

Figure 1. Single Cell CRISPR Screening Workflow



figure2.jpg

Figure 2. Single Cell CRISPR Principles

  • Functional Genomics Research

  • Tumor Mechanism Research

  • Drug Target Screening

  • Immune Regulation Mechanism

  • Stem Cell Regulatory Research

  • Infectious Disease Research


Grouping_plot_by_sgRNA.jpg

Grouping plot by sgRNA

Target_gene_expression_violin_plot.jpg

Target gene expression violin plot


Correlation_heatmap_between_sgRNA_subgroups.jpg

Correlation heatmap between sgRNA subgroups


Dimensionality_reduction_visualization_of_sgRNA_subgroups_by_phenotype.jpg

Dimensionality reduction visualization of sgRNA subgroups by phenotype


Density_and_contour_plots_of_the_expression_of_the_specified_sgRNA_target_genes_in_the_UMAP_plot.jpg

Density and contour plots of the expression of the specified sgRNA target genes in the UMAP plot


Heatmap_of_correlation_between_sgRNA_subgroups_and_gene_expression_subgroups.jpg

Heatmap of correlation between sgRNA subgroups and gene expression subgroups


sgRNA_subgroup_calculation_boxplot_of_a_gene_set_score.jpg

sgRNA subgroup calculation boxplot of a gene set score


Transcriptome_subgroup_marker_gene_heatmap_and_list_of_significantly_associated_sgRNA_target_genes.jpg

Transcriptome subgroup marker gene heatmap and list of significantly associated sgRNA target genes


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