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dHyperCas12a enables multiplexed CRISPRi screens
Schuyler M Melore1,2,3,4, Christian D McRoberts Amador3,4,5, Marisa C Hamilton1,3,4
1University Program in Genetics & Genomics, Duke University, Durham, NC, USA.
Nature Communications
|February 11, 2026
Summary
Researchers developed a new CRISPR tool, dHyperLbCas12a, for precise control over gene and cis-regulatory element activity. This system enables highly multiplexed gene expression studies and manipulation in various cell types.
Area of Science:
- Molecular Biology
- Genetics
- Gene Regulation
Background:
- Gene and cis-regulatory element (CRE) interactions are crucial for biological processes.
- CRISPR screens enable gene/CRE activity assessment, but interaction analysis is limited.
Purpose of the Study:
- To develop a system for efficient, highly multiplexed control of regulatory element activity.
- To enable dissection of independent and combinatorial contributions of CREs to gene expression.
Main Methods:
- Combined a hyper-efficient dCas12a (dHyperLbCas12a) with long CRISPR RNA (crRNA) arrays.
- Utilized RNA Polymerase II for expression of crRNA arrays.
- Applied dHyperLbCas12a for simultaneous activation and repression.
Main Results:
- Demonstrated multiplexed control of gene expression using activation and repression domains.
- Successfully applied the system in cultured primary immune cells and for iPSC differentiation.
- Showcased the ability to dissect independent and combinatorial CRE contributions.
Conclusions:
- The dHyperLbCas12a system offers efficient, highly multiplexed control of gene expression.
- This technology expands possibilities for studying gene regulation in diverse biological systems.
- Enables detailed analysis of cis-regulatory element functions and interactions.
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