• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文学位 >RNA Directed Strategies for the Treatment of HIV-1 Infection.
【24h】

RNA Directed Strategies for the Treatment of HIV-1 Infection.

机译：RNA指导的HIV-1感染治疗策略。

获取原文
获取原文并翻译 | 示例
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Many strategies for HIV-1 treatment are being tested clinically. Since there are still limited options for patients affected by HIV-1, we have proposed two different RNA based strategies to expand these options. We chose RNA directed strategies due to efficacy and advantages in regards to potential cell-type delivery systems and viral vector production.;Our first strategy was to use aptamers targeting HIV-1 gp120 protein to carry mRNA into the cell. The following document provides proof of principle studies delivering reporter mRNA cargo to gp120 presenting cells using the aptamer. Unfortunately, we failed to obtain robust protein expression due to poor serum stability of the mRNA and endosomal trapping upon cellular uptake. Instead of proceeding with aptamer mediated delivery, we switched to liposomal delivery using commercially available lipid reagents. Further proof-of-concept studies were conducted to track mRNA upon systemic delivery and to edit cells in vivo using cre recombinase encoding mRNA. By obtaining recombination in vivo using this approach, we believe that other approaches for gene editing using systemically delivered mRNA are feasible.;Genome editing is a powerful way to obtain a permanent gene deletion, addition, or inversion by the introduction of a molecule capable of DNA cleavage. Multiple forms of genome editing are available; including cre recombinase induced genetic recombination and the clustered regularly interspaced short palindromic repeats (CRISPR)/ cas9 system. To represent a more clinically relevant strategy for HIV-1 treatment, we have used the CRISPR/cas9 system to eliminate HIV viral protein production from cell lines and primary cells with a lentiviral vector delivery. Current limitations in regards to delivery prohibited us from using an mRNA approach as the nucleic acid material encoding gene editing enzymes, but the lentivirus system was extremely successful. Using mRNA is still advantageous to limit potential off-target effects, but further development of delivery systems for nucleic acid delivery of gene editing enzymes is necessary.
机译：目前正在对HIV-1治疗的许多策略进行临床测试。由于受HIV-1影响的患者选择仍然有限，我们提出了两种基于RNA的不同策略来扩展这些选择。由于在潜在细胞类型传递系统和病毒载体生产方面的功效和优势，我们选择了RNA定向策略。我们的第一个策略是使用靶向HIV-1 gp120蛋白的适体将mRNA携带到细胞中。以下文件提供了使用适配子将报告基因mRNA货物转运至gp120呈递细胞的原理研究证据。不幸的是，由于mRNA的较差的血清稳定性和细胞摄取后的内体捕获，我们未能获得稳定的蛋白表达。代替进行适体介导的递送，我们改用市售脂质试剂进行脂质体递送。进行了进一步的概念验证研究，以追踪全身递送后的mRNA并使用编码cre的重组酶mRNA体内编辑细胞。通过使用这种方法在体内获得重组，我们相信使用系统递送的mRNA进行基因编辑的其他方法也是可行的。基因组编辑是一种强大的方法，可以通过引入一种能够将基因永久性删除，添加或倒置的分子来实现。 DNA切割。基因组编辑有多种形式。包括cre重组酶诱导的基因重组和成簇的规则间隔的短回文重复序列（CRISPR）/ cas9系统。为了代表治疗HIV-1的更具临床意义的策略，我们使用了CRISPR / cas9系统，通过慢病毒载体的递送消除了细胞系和原代细胞中HIV病毒蛋白的产生。关于递送的当前限制禁止我们使用mRNA方法作为编码基因编辑酶的核酸材料，但是慢病毒系统非常成功。使用mRNA仍然有利于限制潜在的脱靶作用，但是需要进一步开发用于核酸递送基因编辑酶的递送系统。

著录项

  • 作者

    Bobbin, Maggie.;

  • 作者单位

    City of Hope's Irell & Manella Graduate School of Biomedical Sciences.;

  • 授予单位 City of Hope's Irell & Manella Graduate School of Biomedical Sciences.;
  • 学科 Molecular biology.
  • 学位 Ph.D.
  • 年度 2015
  • 页码 155 p.
  • 总页数 155
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 S-4;
  • 关键词

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号