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Antibody-mediated delivery of CRISPR-Cas9 ribonucleoproteins in human cells

Protein Eng Des Sel. 2022-11; 
Stephanie Ubiparipovic, Daniel Christ, Romain Rouet
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Gene Synthesis … at acidic pH (variant named Pertuzumab-SG), thereby enhancing cytotoxicity of the antibody-… Fab heavy (His-tagged) and light chains were gene synthesized (Genscript) and cloned into … Get A Quote

摘要

The CRISPR genome editing technology holds great clinical potential for the treatment of monogenetic disorders such as sickle cell disease or muscular dystrophy. The therapeutic in vivo application of the technology relies on targeted delivery methods of the Cas9 and gRNA complex to specific cells and/or tissues. However, such methods are currently limited to direct organ delivery, preventing wide clinical application. Here, we show that monoclonal antibodies can be employed to deliver the Cas9/gRNA complex directly into human cells via cell-surface receptors. Using the SpyCatcher/SpyTag system, we conjugated the Fab fragment of the therapeutic antibodies Trastuzumab and Pertuzumab directly to the Cas9 enzyme a... More

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