In Vivo Histone Labeling Using Ultrafast trans-Splicing Inteins.
| Autor: | Prescott NA; Tri-Institutional PhD Program in Chemical Biology, New York, NY, USA.; Chemical Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA., David Y; Tri-Institutional PhD Program in Chemical Biology, New York, NY, USA. davidshy@mskcc.org.; Chemical Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA. davidshy@mskcc.org.; Department of Pharmacology, Weill Cornell Medicine, New York, NY, USA. davidshy@mskcc.org.; Department of Physiology, Biophysics and Systems Biology, Weill Cornell Medicine, New York, NY, USA. davidshy@mskcc.org. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2020; Vol. 2133, pp. 201-219. |
| DOI: | 10.1007/978-1-0716-0434-2_10 |
| Abstrakt: | 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. |
| Databáze: | MEDLINE |
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