CRISPR screens in primary human cells for target identification and validation

Myllia performs large-scale CRISPR screens in primary human cells to identify drug targets using functional genomics. Our approach captures primary human cell biology and thus increases the chance of success for your next drug discovery campaign.

Figure 1: Workflow of CROP-Seq screens in primary human immune cells

Myllia pioneers in primary cell CRISPR screens conducted in human, clinically relevant models such as primary human immune cells that have previously not been accessible for drug target identification at scale. CRISPR-based functional genomics screening has emerged as a powerful tool to unravel factors involved in primary human immune cell function, thus paving the way for novel treatments in oncology, autoimmunity and inflammation.

Briefly, to capture relevant human disease biology, Myllia specializes in high-content CRISPR screens in primary human cell models such as:

T cells (T-lymphocytes)
Myeloid cells (dendritic cells and macrophages)
Epithelial cells

To enable the discovery of novel targets, we have performed multiple CROP-Seq CRISPR KO screens in primary human T cells investigating T cell stemness and effector phenotypes as well as CD4+ T helper cell differentiation towards Th1, Th2, or Th17 subsets. As an example, Myllia has performed a T cell CROP-Seq screen for mediators of primary Th2 cell differentiation (Figure 2).

Th2 cell differentiation

Figure 2: Primary T cell activation and skewing towards the Th2 subset

Using scRNA-Seq in combination with CITE-Seq, we can annotate distinct T helper cell subsets and pinpoint to mRNA transcripts that specify T cell functional plasticity. In this case, Th2 cells can be identified using multiple transcriptomic markers such as GATA3 and IFNGR2 (Figure 3).

Transcriptomic signatures

Figure 3: Transcriptomic signatures define specific T cell subsets

The knockout of IL4R and HDAC3 genes resulted in depletion in differentiated Th2 cells, suggesting their importance in Th2 cell differentiation (Figure 4). On the other hand, the knockout of IL2RA and NFATC2 was found to be enriched in differentiated Th2 cells (Figure 4), indicating their potential role in promoting Th2 cell differentiation.

Gene knockouts

Figure 4: Perturbation effects on primary human Th2 cell differentation

Would you like to take your drug discovery campaign to the next level? If so, please

• Download the introductory slide deck on Myllia’s CRISPR screening platform in primary human cells here
• Reach out to info@myllia.com if you would like to discuss your next CRISPR screening project in primary human cells with us!