In Vivo Histone Labeling Using Ultrafast trans-Splicing Inteins
- 195 Downloads
The development of expressed protein ligation (EPL) widened the scope of questions that could be addressed by mechanistic biochemistry. Protein trans-splicing (PTS) relies on the same basic chemical principles, but utilizes split inteins to tracelessly ligate distinct peptide or polypeptide fragments together with native peptide bonds. Here we present a method to adapt PTS methodologies for their use in live cells, in order to deliver synthetic or native histone modifications. As an example, we provide a protocol to incorporate a small molecule fluorophore into chromatinized histones. The protocol should be easily adaptable to incorporate other modifications to chromatin in vivo.
Key wordsSplit inteins In cellulo labeling Synthetic biology Chromatin
Work in the David lab is supported by the Josie Robertson Foundation, the Pershing Square Sohn Cancer Alliance, Mr. William H. Goodwin and Mrs. Alice Goodwin and the Commonwealth Foundation for Cancer Research and the Center for Experimental Therapeutics at Memorial Sloan Kettering Cancer Center, the CCSG core grant P30 CA008748, and the NIH CEBRA award # DA044767. N.A.P. is supported by the NIH T32 GM115327-Tan chemistry-biology interface training grant and the National Science Foundation Graduate Research Fellowship Grant Number 2017239554.