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ARTTM Barrier Specialty Pipette Tips: 10 µl

Company: Thermo Fisher Scientific
Catalog#: 2140
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Induction of Epithelial-mesenchymal Transition in MDCK II Cells
Author:
Date:
2021-02-05
[Abstract]  

Epithelial-mesenchymal transition (EMT) is a reversible process of epithelial cell transdifferentiation into a mesenchymal cell, that enables initiation of cell migration. EMT plays an important role in embryonic development, tissue repair and cancer metastasis. Better understanding of cellular and molecular events during EMT will not only provide novel insights on how mammalian organism develops and how epithelial tissues regenerate, but also can identify novel therapeutic targets for cancer therapy. Here we aim to provide a detailed protocol on how to induce EMT in Madin-Darby Canine Kidney (MDCK) II epithelial cell line and perform immunofluorescent staining on EMT-induced cells.

[摘要]  [摘要]上皮-间质转化(EMT)是上皮细胞向分化为间充质细胞的可逆过程,能够启动细胞迁移。EMT在胚胎发育,组织修复和癌症转移中起着重要作用。更好地了解EMT过程中的细胞和分子事件,不仅将为哺乳动物生物的发育以及上皮组织的再生提供新颖的见解,而且还可为癌症治疗确定新的治疗靶标。在这里,我们旨在提供有关如何在Madin-Darby犬肾脏(MDCK)II上皮细胞系中诱导EMT并在EMT诱导的细胞上进行免疫荧光染色的详细协议。


[背景]上皮细胞的特征在于细胞可塑性,即具有采用不同细胞表型的能力(Carter等,2019; Yuan等,2019)。上皮-间质转化(EMT)是上皮细胞可塑性的一种形式。在EMT期间,上皮细胞会破坏细胞间连接,使其极性失去作用,并从鳞状,长方体或柱状变为梭形并迁移,从而获得间充质细胞的特性(Kalluri和Weinberg,2009)。EMT可以通过免疫染色和测量标志物(例如E-钙粘蛋白,ZO-1,波形蛋白,纤连蛋白和N-钙粘蛋白)的表达水平进行评估(Kalluri和Weinberg,2009)。在过去十年中在EMT研究中心大多的转录因子的作用例如,SNAIL1 / 2,ZEB1 / ...

Improving CRISPR Gene Editing Efficiency by Proximal dCas9 Targeting
Author:
Date:
2017-08-05
[Abstract]  Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated (Cas) systems function as an adaptive immune system in bacteria and archaea for defense against invading viruses and plasmids (Barrangou and Marraffini, 2014). The effector nucleases from some class 2 CRISPR-Cas systems have been repurposed for heterologous targeting in eukaryotic cells (Jinek et al., 2012; Cong et al., 2013; Mali et al., 2013; Zetsche et al., 2015). However, the genomic environments of eukaryotes are distinctively different from that of prokaryotes in which CRISPR-Cas systems have evolved. Mammalian heterochromatin was found to be a barrier to target DNA access by Streptococcus pyogenes Cas9 (SpCas9), and nucleosomes, the basic units of ... [摘要]  集群定期间隔短回归重复(CRISPR)和CRISPR相关(Cas)系统作为细菌和古菌中的适应性免疫系统,用于防御入侵病毒和质粒(Barrangou和Marraffini,2014)。来自某些2类CRISPR-Cas系统的效应核酸酶已被重新用于真核细胞中的异源靶向(Jinek et al。,2012; Cong等人,2013; Mali ,2013; Zetsche等人,2015)。然而,真核生物的基因组环境与CRISPR-Cas系统发展的原核生物的基因组环境有明显的不同。发现哺乳动物异染色质是通过化脓性链球菌Cas9(SpCas9)靶向DNA接近的障碍,并且还发现染色质的基本单位的核小体阻碍了通过SpCas9的靶DNA进入和切割[ (Knight等人,,2015; Hinz等人,2015; Horlbeck等人,2016年) ; ...

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