Novel epigenome editors for multiplexed gene regulation

Unmet Need

Epigenome editors activate or inhibit gene expression without modifying the underlying DNA sequence. Consequently, these editors can reversibly fine-tune gene expression without the risk of permanently altering the genome or off-target editing. Epigenome editors are currently in development for therapeutic purposes to restore gene function or to silence undesirable gene expression. However, most epigenome editors can only modulate one gene or regulatory region at a time, limiting its ability to achieve combinatorial gene regulation that underlies complex biological processes. There is a need for an epigenome editor that is compatible with multiplexed gene regulation.

Technology

Duke inventors have identified a novel epigenome editing method for multiplexed control of gene regulation, intended for research and clinical applications. The inventors have developed a toolbox of catalytically inactive dHyperLbCas12a-based epigenome editors to guide p300 and SID effectors to targeted regions for gene activation and inhibition. This system supports complex pre-crRNAs, enabling simultaneous activation and repression of multiple genomic regions, enhancing experimental design. The method has been demonstrated to effectively control gene expression in vitro using human cell lines, with lower copy numbers of effectors increasing its efficiency and versatility.

Other Applications

In addition to targeting specific genomic regions, this invention can be used as a screening tool to discover combinatorial interactions of genomic regions underlying a specific phenotype. This tool can also be adapted for use in other organisms (ex. improving yield in plants, increasing resistance to pathogens), enabling potential industrial applications. It also opens avenues for studying complex genetic interactions in health and disease, paving the way for innovative therapeutic strategies.

Advantages

• Utilizes the hyper-efficient dHyperLbCas12a system, enhancing multiplexing capabilities
• Compatible with the activation and inhibition of multiple genomic regions at once
• Supports high-throughput applications for a broader understanding of combinatorial regulatory element activity
• Validated fine-tuning of gene expression in vitro