On-microscope staging of live cells reveals changes in the dynamics of transcriptional bursting during differentiation.
On-microscope staging of live cells reveals changes in the dynamics of transcriptional bursting during differentiation.
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DOI：
10.1038/s41467-022-33977-4
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发表时间：
2022-11-04
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
--
作者：
确定基因在发育过程中开启和关闭的机制是当前生物医学研究的一个关键目标。基因转录已被广泛观察到以不连续的方式发生，短时间的活性爆发穿插着不活动的时期。目前尚不清楚这种动态行为是否或如何随着哺乳动物细胞分化而变化。为了研究这一点，使用显微镜分析，我们在整个红细胞生成过程中监测活细胞中的小鼠α-珠蛋白转录。我们发现，α-珠蛋白转录的总体水平的变化与基因处于活跃转录状态的时间分数的变化最密切相关。我们确定在整个分化的转录爆发模式的差异，最大的转录活性发生在分化的中期。在分化的早期，我们观察到转录活性的波动增加，而在早期成红细胞中，在基因表达的高峰期，转录相对稳定。在分化后期，随着α-珠蛋白表达的下降，我们再次观察到单个细胞内转录的更多变异性。我们建议，所观察到的转录行为的变化可能反映了活性转录区室的稳定性的变化，因为基因表达在分化过程中受到调节。在这里，作者通过结合α-珠蛋白RNA转录物的PP7标记和"显微镜下"细胞分期来观察整个红细胞生成过程中的实时转录动力学。它们显示了在分化过程中转录爆发的模式如何变化，在基因表达的高峰期，单个细胞的变异性显著降低。
Determining the mechanisms by which genes are switched on and off during development is a key aim of current biomedical research. Gene transcription has been widely observed to occur in a discontinuous fashion, with short bursts of activity interspersed with periods of inactivity. It is currently not known if or how this dynamic behaviour changes as mammalian cells differentiate. To investigate this, using an on-microscope analysis, we monitored mouse α-globin transcription in live cells throughout erythropoiesis. We find that changes in the overall levels of α-globin transcription are most closely associated with changes in the fraction of time a gene spends in the active transcriptional state. We identify differences in the patterns of transcriptional bursting throughout differentiation, with maximal transcriptional activity occurring in the mid-phase of differentiation. Early in differentiation, we observe increased fluctuation in transcriptional activity whereas at the peak of gene expression, in early erythroblasts, transcription is relatively stable. Later during differentiation as α-globin expression declines, we again observe more variability in transcription within individual cells. We propose that the observed changes in transcriptional behaviour may reflect changes in the stability of active transcriptional compartments as gene expression is regulated during differentiation. Here the authors observe transcription dynamics in real-time throughout erythropoiesis by combining PP7 tagging of α-globin RNA transcripts and “on-microscope” cell staging. They show how the pattern of transcriptional bursting changes during differentiation, with variability in individual cells significantly reduced at the peak period of gene expression.
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影响因子：
5.3
作者：
KELLER, G;KENNEDY, M;WILES, MV
通讯作者：
WILES, MV
DOI：
10.1073/pnas.1703087114
发表时间：
2017-09-05
影响因子：
11.1
作者：
Jeziorska, Danuta M.;Murray, Robert J. S.;Tufarelli, Cristina
通讯作者：
Tufarelli, Cristina
CD71high 群体代表源自小鼠胚胎干细胞的原始成红细胞
影响因子：
1.2
作者：
Chao, Ruihua;Gong, Xueping;Wang, Yuan
通讯作者：
Wang, Yuan
影响因子：
64.5
作者：
Boija A;Klein IA;Sabari BR;Dall'Agnese A;Coffey EL;Zamudio AV;Li CH;Shrinivas K;Manteiga JC;Hannett NM;Abraham BJ;Afeyan LK;Guo YE;Rimel JK;Fant CB;Schuijers J;Lee TI;Taatjes DJ;Young RA
通讯作者：
Young RA
影响因子：
9.9
作者：
Fritzsch C;Baumgärtner S;Kuban M;Steinshorn D;Reid G;Legewie S
通讯作者：
Legewie S