Ono Shoichiro
Departments of Pathology and Cell Biology, Emory University School of Medicine, Atlanta, GA, United States.
Winship Cancer Institute, Emory University School of Medicine, Atlanta, GA, United States.
Front Cell Dev Biol. 2024 Nov 13;12:1504980. doi: 10.3389/fcell.2024.1504980. eCollection 2024.
Lifeact is a short peptide that is widely utilized as a probe for actin filaments in live imaging. However, high concentrations of Lifeact can alter actin filament dynamics and cause artificial modifications to the actin cytoskeleton. Here, I evaluated strains expressing Lifeact fused to fluorescent proteins in the body wall muscle. I found that, while low-level expression of Lifeact from a single-copy transgene was appropriate for labeling sarcomeric actin filaments, overexpression of Lifeact from an extrachromosomal array causes severe disorganization of muscle sarcomeres and lethality at an embryonic or larval stage. Therefore, for imaging studies in , Lifeact needs to be kept at a low level by proper management of the expression system.
Lifeact是一种短肽,在活细胞成像中被广泛用作肌动蛋白丝的探针。然而,高浓度的Lifeact会改变肌动蛋白丝的动力学,并对肌动蛋白细胞骨架造成人为修饰。在这里,我评估了在体壁肌肉中表达与荧光蛋白融合的Lifeact的菌株。我发现,虽然单拷贝转基因低水平表达Lifeact适用于标记肌节肌动蛋白丝,但染色体外阵列过量表达Lifeact会导致肌肉肌节严重紊乱,并在胚胎或幼虫阶段致死。因此,对于秀丽隐杆线虫中的成像研究,需要通过适当管理表达系统将Lifeact保持在低水平。