Design of Synthetic Combinatorial Transcription Factors

描述（由申请人提供）：许多生物决定是由复杂的转录电路控制的。可以特异性调节这种回路的试剂将是非常有用的，作为解剖复杂过程的顺式调节逻辑的工具，并最终作为治疗剂。最好的开发工具是合成的DNA结合蛋白，但这些缺乏在天然转录因子中观察到的组合特异性。最终，通过一系列协调一致的设计因素，可以实现更精确的监管。许多天然的组合转录因子充当变构开关-该因子仅在另一辅因子存在时才有活性。最近，已经开发了一种用于引入对蛋白质活性的变构控制的新策略。该策略涉及将构象自抑制相互作用引入蛋白质中;通过与适当的伴侣蛋白相结合可以减轻光抑制。我的具体目标是使用Ithis自抑制策略来产生合成的转录因子，其中：1）DNA结合活性由邻近的DNA结合辅因子变构调节。2)转录激活功能由邻近的DNA结合辅因子变构调节。这些设计因子应该产生非常紧密的转录控制-激活依赖于分子的精确组合的同时存在。

DESCRIPTION (provided by applicant): Many biological decisions are controlled by complex transcriptional circuits. Reagents that can specifically modulate such circuits would be extremely useful, as tools for dissecting the cis-regulatory logic of complex processes, and ultimately as therapeutics. The best developed tools are synthetic DNA binding proteins, but these lack the combinatorial specificity observed in natural transcription factors. Ultimately, more precise regulation could be achieved with sets of designer factors that act in concert. Many natural combinatorial transcription factors act as allosteric switches -- the factor is only active in the presence of another co-factor. Recently a novel strategy for introducing allosteric control over a protein activity has been developed. This strategy involves the introduction of conformational autoinhibitory interactions into a protein; lautoinhibition can be relieved by association with an appropriate partner protein. My specific aims are to use Ithis autoinhibitory strategy to generate synthetic transcription factors in which: 1) DNA binding activity is allosterically regulated by an adjacent DNA binding co-factor. 2) Transcriptional activation function is allosterically regulated by an adjacent DNA binding co-factor. These designer factors should yield extremely tight transcriptional control -- activation is dependent on the simultaneous presence of a precise combination of molecules.