KLF4 N-terminal variance modulates induced reprogramming to pluripotency.
KLF4 N-terminal variance modulates induced reprogramming to pluripotency.
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DOI：
10.1016/j.stemcr.2015.02.004
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发表时间：
2015-04-14
影响因子：
5.9
通讯作者：
Woltjen, Knut
中科院分区：
文献类型：
--
作者：
Kim, Shin-Il;Oceguera-Yanez, Fabian;Hirohata, Ryoko;Linker, Sara;Okita, Keisuke;Yamada, Yasuhiro;Yamamoto, Takuya;Yamanaka, Shinya;Woltjen, Knut
作为经典的重编程模型，Oct3/4、SOX2、KLF4和c-myc重新连接体细胞以实现诱导多能性。然而，方法上的细微差异混淆了对重新编程机制的比较研究。利用转座子，我们系统地评估了不同多顺反子盒对小鼠体细胞重新编程的细胞和分子特征。重编程反应在转基因非依赖性多能性的启动和稳定程度上有所不同。值得注意的是，这些盒式磁带使用了两个KLF4变体中的一个，只有九个N末端氨基酸不同，这产生了不同的蛋白质化学计量比。将较短的变体延长9个N-末端氨基酸或通过补充KLF4增加化学计量比既挽救了蛋白质水平，又挽救了表型差异，暗示了决定重新编程结果的阈值。引人注目的是，已发表的多顺反子盒式磁带引发的全球基因表达模式因KLF4变体的不同而不同。我们的数据揭示了KLF4参考cDNA变异，它改变了多顺反子因子的化学计量学，预测了重新编程的标志，并指导了兼容的公共数据集的比较。重编程载体不一致地使用两个未被识别的KLF4变体中的一个编码KLF4 N末端变体编码多顺反子盒驱动不同的化学计量比重编程启动和稳定对KLF4蛋白水平敏感因此，由公共载体诱导的基因表达形成两个不同的簇在本文中，Woltjen和他的同事揭示了不知情地使用两个不同的KLF4 cDNA来构建体细胞重编程载体。克隆在多顺反子盒式磁带中，每个变体产生不同水平的KLF4蛋白，影响重新编程表型。此外，IPSC衍生过程中的比较基因表达基于所使用的KLF4 cDNA而分叉，这支持了因子化学计量学在定义重编程结果中的重要性。
As the quintessential reprogramming model, OCT3/4, SOX2, KLF4, and c-MYC re-wire somatic cells to achieve induced pluripotency. Yet, subtle differences in methodology confound comparative studies of reprogramming mechanisms. Employing transposons, we systematically assessed cellular and molecular hallmarks of mouse somatic cell reprogramming by various polycistronic cassettes. Reprogramming responses varied in the extent of initiation and stabilization of transgene-independent pluripotency. Notably, the cassettes employed one of two KLF4 variants, differing only by nine N-terminal amino acids, which generated dissimilar protein stoichiometry. Extending the shorter variant by nine N-terminal amino acids or augmenting stoichiometry by KLF4 supplementation rescued both protein levels and phenotypic disparities, implicating a threshold in determining reprogramming outcomes. Strikingly, global gene expression patterns elicited by published polycistronic cassettes diverged according to each KLF4 variant. Our data expose a Klf4 reference cDNA variation that alters polycistronic factor stoichiometry, predicts reprogramming hallmarks, and guides comparison of compatible public data sets. Reprogramming vectors inconsistently employ one of two unappreciated Klf4 variants Polycistronic cassettes encoding Klf4 N-terminal variants drive distinct stoichiometry Reprogramming initiation and stabilization are sensitive to Klf4 protein levels Accordingly, gene expression elicited by public vectors forms two distinct clusters In this article, Woltjen and colleagues reveal the unwitting employment of two different Klf4 cDNAs to construct somatic cell reprogramming vectors. Cloned in a polycistronic cassette, each variant produces different KLF4 protein levels, impacting reprogramming phenotypes. Moreover, comparative gene expression during iPSC derivation bifurcates based on the Klf4 cDNA used, underpinning the importance of factor stoichiometry in defining reprogramming outcomes.
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影响因子：
64.8
作者：
Israel, Mason A.;Yuan, Shauna H.;Bardy, Cedric;Reyna, Sol M.;Mu, Yangling;Herrera, Cheryl;Hefferan, Michael P.;Van Gorp, Sebastiaan;Nazor, Kristopher L.;Boscolo, Francesca S.;Carson, Christian T.;Laurent, Louise C.;Marsala, Martin;Gage, Fred H.;Remes, Anne M.;Koo, Edward H.;Goldstein, Lawrence S. B.
通讯作者：
Goldstein, Lawrence S. B.
影响因子：
4.6
作者：
Guo, Ge;Yang, Jian;Smith, Austin
通讯作者：
Smith, Austin
DOI：
10.1073/pnas.1004584107
发表时间：
2010-05-18
影响因子：
11.1
作者：
Hanna, Jacob;Cheng, Albert W.;Jaenisch, Rudolf
通讯作者：
Jaenisch, Rudolf
DOI：
10.1073/pnas.0711983105
发表时间：
2008-02-26
影响因子：
11.1
作者：
Lowry, W. E.;Richter, L.;Plath, K.
通讯作者：
Plath, K.
影响因子：
23.9
作者：
Carey, Bryce W.;Markoulaki, Styliani;Jaenisch, Rudolf
通讯作者：
Jaenisch, Rudolf