Department of Chemistry and Biochemistry, University of California, Santa Barbara, CA, 93106, USA.
Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, MN, 55455, USA.
Small. 2018 Jul;14(30):e1800543. doi: 10.1002/smll.201800543. Epub 2018 Jul 2.
A light-activated genome editing platform based on the release of enzymes from a plasmonic nanoparticle carrier when exposed to biocompatible near-infrared light pulses is described. The platform relies on the robust affinity of polyhistidine tags to nitrilotriacetic acid in the presence of copper which is attached to double-stranded nucleic acids self-assembled on the gold nanoparticle surface. A protein fusion of the Cre recombinase containing a TAT internalization peptide sequence to achieve endosomal localization is also employed. High-resolution gene knock-in of a red fluorescent reporter is observed using a commercial two-photon microscope. High-throughput irradiation is described to generate useful quantities of edited cells.
描述了一种基于光激活的基因组编辑平台,该平台基于等离子体纳米粒子载体在暴露于生物相容性近红外光脉冲时释放酶。该平台依赖于多组氨酸标签在铜存在下与氮三乙酸的牢固亲和力,铜附着在自组装在金纳米粒子表面的双链核酸上。还使用了 Cre 重组酶的蛋白质融合,其中包含 TAT 内化肽序列以实现内体定位。使用商业双光子显微镜观察到红色荧光报告基因的高分辨率基因敲入。还描述了高通量辐照以产生有用数量的编辑细胞。