DESIGNER DNA BINDING PROTEINS TARGETING HIV GENES

锌指是一类DNA结合蛋白， 根据最近设计的识别方法来识别DNA序列 代码.这一发展开辟了设计 嵌合转录抑制因子、反式激活因子和核酸内切酶 用于调节或灭活特定的病毒和人类基因。我们 我建议利用这项新兴技术来开发能够 抑制HIV必需基因的表达。在第一阶段，我们 构造并表征了三个设计者的结合亲和力 锌指蛋白，靶向邻近保守的9个碱基对 在第二阶段，我们将确定 这些蛋白质结合HIV DNA序列并减弱生长的能力 体内的艾滋病病毒。我们还将锌指蛋白对与 肽接头，以构建具有多达18个 碱基对特异性这项研究将导致发展 用于减弱病毒繁殖的治疗药物。更一般地说， 设计DNA结合蛋白技术的发展将导致 用于治疗多种病毒性疾病的新疗法的开发 和人类疾病，包括癌症、传染病和自身免疫性疾病， 紊乱 拟定商业应用： 我们建议开发的设计师DNA结合蛋白将是一种 这是一种新型抗病毒药物的重要原理证明。他们的 最直接的潜在应用是检测病毒DNA 诊断建议的序列。长期潜在治疗 应用是选择性地抑制病毒基因表达， 感染的细胞，以帮助减少病毒载量。

Zinc fingers are a class of DNA binding proteins that can be designed to recognize DNA sequences according to a recently devised recognition code. This development has opened up the possibility of designing chimeric transcription respressors, transactivators and endonucleases for regulating or inactivating specific viral and human genes. we propose to utilize this emerging technology to develop drugs capable of inhibiting expression of an essential HIV gene. In Phase I, we constructed and characterized the binding affinities for three designer zinc finger proteins that target adjacent conserved, 9 base-pair sequences in an essential HIV gen. In Phase II, we will determine the ability of these proteins to bind HIV DNA sequences and attenuate growth of HIV in vivo. We will also link pairs of the zinc finger proteins with a peptide linker to construct hybrid DNA binding proteins with up to 18 base-pair specificity. This research will lead to the development of therapeutic drugs for attenuating viral reproduction. More generally, the development of designer DNA binding protein technology will lead to the development of novel therapeutics for treating a variety of viral and human diseases, including cancer, infectious diseases and autoimmune disorders. PROPOSED COMMERCIAL APPLICATION: The designer DNA binding proteins that we propose to develop will be an important proof-of-principle for a new class of anti-viral drugs. Their most immediate potential application is in the detection of viral DNA sequences for diagnostic proposes. the longer term potential therapeutic applications are to selectively inhibit viral gene expression within infected cells to help in reducing viral load.