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Direct cytosolic delivery of proteins and CRISPR-Cas9 genome editing by gemini amphiphiles via non-endocytic translocation pathways

Le Z, Pan Q, He A, Liu H, Shi Y, Liu L, Liu Z, Ping Y, Chen Y (2023) Direct cytosolic delivery of proteins and CRISPR-Cas9 genome editing by gemini amphiphiles via non-endocytic translocation pathways. ACS Cent Sci 9(7):1313-1326. doi: 10.1021/acscentsci.3c00207 PMID: 37521791

Objective: Create a library composed of 150 gemini amphiphiles (GAs), a molecule with polar and nonpolar regions, to identify means to facilitate delivery of saporin across the cell membrane.

Summary: The gemini amphiphiles studied differ in the spacing of functional groups, hydrophobicity and sterics. A Cas9 ribonucleoprotein conjugated to saporin showed highest efficacy for internalization. HeLa cells were used to study the efffects of this found gemini-amphiphile conjugated to saporin.

Usage: A1I2-1R2C18 [a gemini ampiphile] conjugated to saporin and treated against HeLa cells at 0.0625, 0.125, 0.25, 0.5, 1μg/mL for 4 hours and then assessed for cell viability.

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