Versatile and robust epigenetic modulation platform for programmable gene activation and repression

Hi, I’m Ryan Rickels, and I’ll be sharing how Elevate Bio’s RNA-guided epigenetic editing platform can help partners regulate disease-relevant genes without changing the underlying DNA sequence, especially when a program requires tunable activation, durable repression, or coordinated control of multiple targets.

That matters because many diseases are driven by abnormally high gene expression or abnormal gene silencing. With epigenetic editing, the goal is to tune gene activity up or down in a precise programable way. On the repression side, we use RNA guided epigenetic repressors to modify the local chromatin environment and reduce transcription of a target gene.

In this poster, we show strong repression of B2M across multiple human cell types, including HEK293T cells, Jurkat cells, and primary human T cells. Importantly, after transient mRNA delivery, B2M and repression remain durable for at least four weeks in Jurkat cells. We also demonstrate robust gene activation by recruiting activation machinery to endogenous genes, our epi-activators can boost expression from the native promoter.

For example, we show strong CD25 activation in Jurkat cells, activation of a metabolic gene target in mouse liver cells, and tunable activation of neurodevelopmental genes in Neuro2a cells. This ability to dial expression up to a desired level could be especially important in diseases where restoring even a twofold increase in gene expression may have therapeutic relevance.

Another advantage of our platform is flexibility. Because our collection includes distinct RNA guided DNA binding systems, we can target multiple genes in the same cell. Here, we demonstrate multiplexed editing by activating CD25 while simultaneously repressing B2M in Jurkat cells. Specificity is also central to the platform. Using RNA seq, we evaluated whether our editors caused unintended changes in gene expression.

In these studies, the intended target was the dominant signal with little to no detectable off target activity. Finally, we show delivery optionality in addition to mRNA delivery. Our compact epigenetic activators can be packaged into all-in-one AAV vectors using CD25 activation as a readout, we demonstrate functional AAV delivery in Jurkat cells. Together, these data support a versatile epigenetic modulation platform for programable gene activation and repression with tunable expression, durable effects, multiplexing potential, high on target specificity, and multiple delivery options.

By demonstrating durable repression, tunable activation, multiplexed regulation on target specificity and delivery flexibility, this platform gives partners a broader set of tools for designing gene modulation strategies around the biology of their target.