Virus-like particles allow precise gene editing and combined CRISPR tests in human immune cells

A virus-like particle (VLP)-based toolkit enables efficient CRISPR genome editing in human myeloid cells.

• The toolkit delivers diverse CRISPR editing methods to human monocytes, macrophages, and dendritic cells while maintaining cell viability.
• VLP-mediated delivery supports gene knockout, base editing, and epigenetic silencing, as well as integration of large DNA sequences with AAV donors.
• Pooled loss-of-function screens in human macrophages identified TNFAIP3 as a central regulator influencing TNF production and CD80 expression.
• Ablation of TNFAIP3 led to a pro-inflammatory cell state resistant to suppressive signals and increased the cytotoxicity of engineered HER2 CAR-macrophages.
• This technology may facilitate unbiased discovery of functional gene targets in primary human myeloid cells.

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