Technologies

The discovery of CRISPR-Cas9 mediated genome editing has created the opportunity to manipulate the genome in many different ways. These technologies allow for the modification of genes, the introduction of fluorescent tags or degrons, and the ability to activate or suppress gene transcription. ScreeninC provides access to expertise, technology platforms and service to implement these technologies in research projects.

1. CRISPR knockout

The CRISPR genome-editing technologies are based on an antiviral defense system that operates in bacteria. This biological system can be utilized in, for example, human cells to target a gene of interest by expressing a gRNA with homology to that gene’s sequence.

2. CRISPR inhibition / activation

CRISPR interference or inhibition (CRISPRi) is a technology that allows for the sequence specific repression of gene expression. By making use of a mutant Cas9 (dCas9) fused to a strong transcriptional repressor domain in combination with an sgRNA targeting the promoter region of the gene, gene-specific repression can be achieved

3. RNA Interference

Prior to the development of CRISPR/Cas9 genome-editing technologies, RNA interference (RNAi) was used broadly to suppress the expression of genes of interest and to screen large gene sets for their role in biological processes and phenotypes.

4. Arrayed and pooled screening

Functional genomic screens can be divided into arrayed and pooled formats. In arrayed screens, each gene is perturbed in an individual well containing one or more cells. In contrast, in pooled screens, sets of genes are perturbed individually in a population of cells that is propagated in large culture flasks or dishes.

5. Equipment

Our expertise and collection of tools is the result of many years of research in which we developed or implemented technologies for model generation and large-scale genomic and small molecule screening. We provide researchers throughout the Netherlands access to this broad range of technologies.

6. CRISPR and RNAi libraries

In the course of two decades, we have performed numerous RNAi and CRISPR screens in both human and mouse cell line models. We were the first to generate a whole-genome, pooled shRNA library and have extensive expertise in the design and generation of custom CRISPR or RNAi pooled libraries.

7. High-content imaging

High-content imaging involves automated image acquisition and analysis of multiparametric data at a single-cell level allowing researchers to capture and analyze a wide range of cellular phenotypes simultaneously.

8. CRISPR genome editing

The goal of CRISPR genome editing is to generate a cell line or animal model with one or more specific genetic modifications such as a gene knockout, introduction of a specific mutation, or knock-in of a gene or other sequence.

9. iPSC models

The generation of induced pluripotent stem cell (iPSC) models involves reprogramming of somatic cells into a pluripotent state while retaining the genetic background. The iPSC models can then be differentiated into specific cell types to, for example, study cellular differentiation or cell type specific biology.