Barker Brooke, Cannon Chloe, Umphlett Hannah, Kim Eun Suk, Keiper Brett D
Biochemistry & Molecular Biology, Brody School of Medicine, East Carolina University, Greenville, North Carolina, United States.
MicroPubl Biol. 2025 Feb 18;2025. doi: 10.17912/micropub.biology.001452. eCollection 2025.
Cellular identity and fate are determined by the proteins synthesized. Initiation of mRNA translation requires an important translation factor, eIF4G ( in ). Embryos use mRNA translational control for spatial and temporal regulation of protein synthesis. Using CRISPR engineering, we added in-frame epitope and fluorescent tags (V5, Myc, Flag, GFP, and mCherry) to IFG-1 . Tagged forms containing the V5 epitope caused embryonic arrest. Internal disruption of the V5 tag restored viability at 25°C. This study demonstrates that the molecular nature of a small epitope tag is sufficient to disrupt embryogenesis.
细胞身份和命运由合成的蛋白质决定。mRNA翻译的起始需要一种重要的翻译因子，即真核起始因子4G（eIF4G）。胚胎利用mRNA翻译控制来进行蛋白质合成的时空调节。我们通过CRISPR基因编辑技术，将框内表位和荧光标签（V5、Myc、Flag、GFP和mCherry）添加到IFG-1上。含有V5表位的标记形式导致胚胎发育停滞。V5标签的内部破坏在25°C时恢复了活力。这项研究表明，一个小表位标签的分子性质足以破坏胚胎发育。
