Chromatin and non-chromatin immunoprecipitations to capture protein-protein and protein-nucleic acid interactions in living cells

Proteins are expressed from genes via sequential biological processes of transcription, mRNA processing, export and translation, and play their roles in maintaining cellular functions via interactions with proteins, DNAs or RNAs. Thus, it is important to study the protein interactions during biological processes in living cells towards understanding their mechanisms-of-action in real time. Methodologies have been developed over the years to study protein interactions in vivo. One state-of-the-art approach is formaldehyde crosslinking-based immuno- or chemi-precipitation to analyze selective as well as genome/proteome-wide interactions in living cells. It is a popular and widely used methodology for cellular analysis of the protein-protein and protein-nucleic acid interactions. Here, we describe this approach to analyze protein-protein/nucleic acid interactions in vivo.

Automated summary

Proteins are expressed through transcription, mRNA processing, export and translation, and their interactions with other proteins, DNAs or RNAs play a crucial role in maintaining cellular functions. Studying protein interactions in living cells is important for understanding their mechanisms-of-action in real time. One state-of-the-art approach is formaldehyde crosslinking-based immuno- or chemi-precipitation, which is widely used for analyzing protein-protein and protein-nucleic acid interactions in cellular analysis.