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Expanding the specificity of DNA targeting by harnessing cooperative assembly

机译:利用协同装配扩大DNA靶向的特异性

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The precise control of developmental and regulatory processes in the cell requires accurate recognition of specific DNA sites. For genomes as large as that of humans, single-molecule-DNA binders have difficulties accurately and specifically recognizing the intended targets. Natural transcription factors overcome these difficulties by forming non-covalent complexes on the DNA with other transcription factors. These cooperative complexes overcome the difficulties of single-molecule transcription factors, allowing specific, combinatorial control of a range of transcriptional targets. Artificial transcription factors have been designed to take advantage of this technique of cooperative assembly, facilitating future studies in whole genome targeting. In contrast to a simple model of component independence, cooperative complexes as a whole often display slightly altered DNA specificity from what would be expected from the analysis of their separate components. The true sequence specificity of cooperative complexes, and thus their presumed in vivo targets, have to be experimentally probed. A number of techniques, such as the cognate site identity array, now allow for rapid, high-throughput determination of the specificity of cooperative complexes.
机译:细胞中发育和调控过程的精确控制要求对特定DNA位点的准确识别。对于与人类一样大的基因组,单分子DNA结合剂难以准确,特异地识别预期目标。天然转录因子通过与其他转录因子在DNA上形成非共价复合物而克服了这些困难。这些合作复合物克服了单分子转录因子的困难,从而可以实现一系列转录靶标的特异性,组合控制。已经设计了人工转录因子以利用这种协同装配技术,从而促进了将来对全基因组靶向的研究。与简单的成分独立性模型相反,协作复合物作为一个整体,通常显示出比其单独成分分析所期望的DNA特异性稍有改变的DNA特异性。合作复合物的真实序列特异性,以及因此推测的它们在体内的靶点,必须通过实验进行探测。现在,许多技术(例如同源位点识别阵列)可以快速,高通量确定合作复合物的特异性。

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