Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142,USA.
The Eugene Bell Center for Regenerative Biology and Tissue Engineering, Marine Biological Laboratory, 7 MBL Street, Woods Hole, MA 02543-1015, USA.
Development. 2024 Oct 15;151(20). doi: 10.1242/dev.202857. Epub 2024 Sep 14.
Delivering molecular tools into oocytes is essential for developmental and reproductive biology. Microinjection, the conventional method, is equipment intensive, often technically challenging and has a low yield, and is impractical in species with delicate oocytes or restricted spawning seasons. To overcome these limitations, we developed VitelloTag, a cost-effective, high-throughput system using vitellogenin-derived fusion proteins to enable efficient cargo delivery via receptor-mediated endocytosis. We demonstrate its utility by delivering Cas9/sgRNA complexes in two distantly related species for gene knockout.
将分子工具递送入卵母细胞对于发育和生殖生物学至关重要。微注射是传统方法，需要大量设备，技术上常常具有挑战性，且产量低，在卵母细胞脆弱或繁殖季节有限的物种中不切实际。为了克服这些限制，我们开发了 VitelloTag，这是一种经济高效、高通量的系统，使用卵黄蛋白原衍生的融合蛋白通过受体介导的内吞作用来实现有效货物传递。我们通过在两个远缘物种中递 Cas9/sgRNA 复合物来证明其用途，以实现基因敲除。
