Human FAP Stable Cell Line - HEK293T

Increasing data reveals that well-regulated RNA localization in living cells is critical for cell survival. The researchers created FAP-seq, a unique RNA proximity labeling technology activated by near-infrared (NIR) light. This approach employs a genetically encoded fluorogen activating protein (FAP) that specifically binds malachite green (MG) derivatives. When MG-HI binds, it produces singlet oxygen, making it possible to label RNA and protein in living cells without the need for washing procedures. FAP-seq enables high-resolution RNA localization studies in complicated biological systems, revealing new details about transcriptome dynamics. Molecular dynamics simulations revealed the structural arrangement of FAP-MG-HI, which improved knowledge of its functionality. Overall, FAP-seq is an effective technique for studying RNA-related biological processes with spatial and temporal precision.

Figure 1. The researchers tested live cell protein labeling using Azide-Fluor 545 conjugation via CuAAC in the Human FAP Stable Cell Line - HEK293T. Western blot validation of protein fractionation using the Thermo Scientific Mem-PER Plus Membrane Protein Extraction Kit. The effectiveness of live cell RNA labeling was tested by RNA dot blot analysis, which revealed dosage dependence with Azide-biotin conjugation after RNA extraction. The relative RNA enrichment in Nucleus-FAP and ER-FAP showed substantial differences. (Li L, et al., 2024)