A new study from the Innovative Genomics Institute (IGI) at UC Berkeley, published in The CRISPR Journal (April 2025), introduces a novel strategy to improve CRISPR-Cas9 gene editing efficiency in primary human immune cells, a key step toward therapeutic applications. Led by Dr. Ross Wilson, the research team developed hairpin internal nuclear localization signals (hiNLS) that are embedded within the Cas9 protein, rather than fused to its ends—a common approach in most constructs.
This innovation enhances nuclear delivery of Cas9 when delivered as a ribonucleoprotein (RNP), which is crucial for therapeutic gene editing due to the enzyme’s short half-life and need for rapid nuclear access. The study evaluated two delivery methods—electroporation and a peptide-enabled, nonviral approach—and found that hiNLS constructs improved editing efficiency across multiple gene targets, including B2M and TRAC, which are important in immune function and T cell therapies.
Importantly, these hiNLS-enhanced Cas9 variants not only increase editing performance but are also scalable and manufacturable at high purity and yield. Wilson’s team is now exploring their potential for self-delivery into clinically relevant cells—building on insights from prior work with CRISPR pioneer Jennifer Doudna—and sees broad translational potential in gene therapy.
Source:
https://www.genengnews.com/topics/genome-editing/new-strategy-enhances-crispr-editing-efficiency-for-therapeutic-use/
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