Epigenetic silencing directs expression heterogeneity of stably integrated multi-transcript unit genetic circuits.
Epigenetic silencing directs expression heterogeneity of stably integrated multi-transcript unit genetic circuits.
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DOI：
10.1038/s41598-021-81975-1
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发表时间：
2021-01-28
影响因子：
4.6
通讯作者：
Zhao W
中科院分区：
文献类型：
--
作者：
Zimak J;Wagoner ZW;Nelson N;Waechtler B;Schlosser H;Kopecky M;Wu J;Zhao W
我们报告说，表观遗传沉默导致使用CRISPR-Cas9整合的多转录单位结构的功能丧失。使用带有选择标记的模块化双色报告系统，我们证明了表达的异质性与序列改变无关，而是与染色体的可及性相关。我们通过甲基化和组蛋白脱乙酰化的小分子抑制剂部分逆转了这种表观遗传沉默。然后，我们通过使用ATAC-seq将每个异质表达的表型与其预期的表观遗传状态相关联。选择性压力增强了每种表达表型的稳定性，这表明整合后持续一个月的表观遗传重塑可以发生。总而言之，我们的数据表明，表观遗传沉默限制了通过双链修复途径整合的多转录单位结构的用途。我们的研究表明，哺乳动物合成生物学家在设计复杂的遗传电路时，应该考虑局部的表观遗传结果。
We report that epigenetic silencing causes the loss of function of multi-transcript unit constructs that are integrated using CRISPR-Cas9. Using a modular two color reporter system flanked by selection markers, we demonstrate that expression heterogeneity does not correlate with sequence alteration but instead correlates with chromosomal accessibility. We partially reverse this epigenetic silencing via small-molecule inhibitors of methylation and histone deacetylation. We then correlate each heterogeneously-expressing phenotype with its expected epigenetic state by employing ATAC-seq. The stability of each expression phenotype is reinforced by selective pressure, which indicates that ongoing epigenetic remodeling can occur for over one month after integration. Collectively, our data suggests that epigenetic silencing limits the utility of multi-transcript unit constructs that are integrated via double-strand repair pathways. Our research implies that mammalian synthetic biologists should consider localized epigenetic outcomes when designing complex genetic circuits.
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在单细胞水平上表观遗传调节的动力学。
DOI：
10.1126/science.aab2956
发表时间：
2016-02-12
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Bintu L;Yong J;Antebi YE;McCue K;Kazuki Y;Uno N;Oshimura M;Elowitz MB
通讯作者：
Elowitz MB
影响因子：
46.9
作者：
Hockemeyer, Dirk;Soldner, Frank;Beard, Caroline;Gao, Qing;Mitalipova, Maisam;DeKelver, Russell C.;Katibah, George E.;Amora, Ranier;Boydston, Elizabeth A.;Zeitler, Bryan;Meng, Xiangdong;Miller, Jeffrey C.;Zhang, Lei;Rebar, Edward J.;Gregory, Philip D.;Urnov, Fyodor D.;Jaenisch, Rudolf
通讯作者：
Jaenisch, Rudolf
DOI：
10.1073/pnas.94.2.575
发表时间：
1997-01-21
影响因子：
11.1
作者：
Chung, JH;Bell, AC;Felsenfeld, G
通讯作者：
Felsenfeld, G
影响因子：
5.9
作者：
Ordovás L;Boon R;Pistoni M;Chen Y;Wolfs E;Guo W;Sambathkumar R;Bobis-Wozowicz S;Helsen N;Vanhove J;Berckmans P;Cai Q;Vanuytsel K;Eggermont K;Vanslembrouck V;Schmidt BZ;Raitano S;Van Den Bosch L;Nahmias Y;Cathomen T;Struys T;Verfaillie CM
通讯作者：
Verfaillie CM
影响因子：
5.9
作者：
Kao, Tim;Labonne, Tanya;Niclis, Jonathan C.;Chaurasia, Ritu;Lokmic, Zerina;Qian, Elizabeth;Bruveris, Freya F.;Howden, Sara E.;Motazedian, Ali;Schiesser, Jacqueline V.;Costa, Magdaline;Sourris, Koula;Ng, Elizabeth;Anderson, David;Giudice, Antonietta;Farlie, Peter;Cheung, Michael;Lamande, Shireen R.;Penington, Anthony J.;Parish, Clare L.;Thomson, Lachlan H.;Rafii, Arash;Elliott, David A.;Elefanty, Andrew G.;Stanley, Edouard G.
通讯作者：
Stanley, Edouard G.