Exp Cell Res 2020 Nov 15;3962:112296. doi: 10.1016/j.yexcr.2020.112296.

Injected cells provide a valuable complement to cell-free systems for analysis of gene expression.

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The aim of this short review is to comment on the advantages of injecting purified molecules into a normal living cell as a complement to the constitution of a cell-free system for analyzing the function of cell components. We emphasize here that the major difference is that, by injection, most components of a cell are maintained at their normal concentration, which is difficult, even if at all possible, to achieve in a cell-free system. We exemplify the benefits of a cell injection system by the efficiency and long duration of DNA transcription by RNA polymerase II, as used by most genes, and by the widespread success of injecting purified messenger RNA for protein synthesis. The most recent work using cell injection also gives a new understanding of a long lasting transcription factor residence on its DNA or chromatin not shown by other procedures. Lastly, we re-visit an old idea that transcription factors that guide cell fate may be stably bound to DNA or chromatin, except at S-phase or mitosis in the cell cycle, when they can undergo exchange with equivalent molecules in the cell.

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