In Vitro Transcriptional Analyses of the Human Globin Genes in the K562 Cell Line
Banks, Crystal M.
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The K562 cell line is a human leukemic cell line that displays erythroid-like characteristics. The embryonic and fetal globin genes are expressed in this cell line, but adult hemoglobin (a2 B2) is not. I was interested in establishing and in vitro transcription system that would selectively transcribe the human globin genes in a manner consistent with that observed in vitro. Selective globin gene transcription was not observed when the cloned K562 B-globin gene was transcribed in vitro using the homologous whole cell extract. In contrast, differential transcription of the o-globin gene and the P-globin gene was observed when K562 chromatin was used as a template in the K562 extract. Using Sl analysis, I was able to detect RNA synthesized from the B-globin gene. There was, however, no detectable B-globin RNA, indicating the lack of B-globin transcription. Thus, selective transcription of fetal and adult genes can be demonstrated in vitro when chromatin is used as a template.