Novel Technologies for intracellular manipulation

 描述(申请人提供)：该SBIR项目旨在开发用于研究、诊断和最终治疗目的的活的真核细胞内部的操纵和标记的新技术。穿透细胞的多肽提供了诱人的前景，可以通过真核细胞的质膜运送广泛的用户定义的蛋白质货物。在可传递生物分子的多样性、蛋白质表达/传递的快速性以及随着时间的推移对蛋白质水平的控制方面，通过CPPS传递货物蛋白具有许多优于转染法的优点。目前的CPP技术要么繁琐，需要生产单独标记的蛋白质，要么缺乏特异性和低效率。我们创新的核心技术是基于细胞穿透性多肽偶联接头蛋白，允许自发装载几乎任何所需的货物蛋白，以便快速输送到细胞中。我们的CPP适配器技术不仅允许交付更多的货物，而且与当前技术相比，在速度和易用性以及安全增强方面具有优势。此外，货物从细胞质和系统的适配器中释放，并可以作为模块试剂盒生产，允许使用CPP-适配器识别的相同标签进行亲和货物蛋白纯化。我们寻求开发的其他功能将允许基于不同CPP的属性(例如，核、线粒体或溶酶体定位)的亚细胞递送，并允许将多个货物同时递送到不同的隔间。

 DESCRIPTION (provided by applicant): This SBIR project seeks to develop novel technologies for the manipulation and labeling of the interior of living eukaryotic cells for research, diagnostic and eventually therapeutic purposes. Cell-penetrating peptides offer the tantalizing prospect of delivering a broad palette of user-defined protein cargoes through the plasma membranes of eukaryotic cells. Cargo protein delivery via CPPs has many advantages over transfection with respect to variety of deliverable biomolecules, rapidity of protein expression/delivery and control of protein levels over time. Current CPP technologies are either cumbersome and require the production of individually labeled proteins, or suffer from a lack of specificity and low efficiency. Our innovative core technology is based on cell-penetrating peptide-coupled adaptor proteins that allow the spontaneous loading of almost any desired cargo protein for rapid delivery into cells. Our CPP-adaptor technology not only allows delivery of an extended array of cargoes, but has advantages in speed and ease of use as well as safety enhancements compared to current technologies. In addition, cargoes are released from the adaptors in the cytoplasm and the system and can be produced as a modular kit allowing affinity cargo protein purification using the same tag recognized by the CPP-adaptor. Additional features we seek to develop will allow subcellular delivery based on the properties of different CPPs (e.g. nuclear, mitochondrial or lysosomal localization) and enable simultaneous delivery of multiple cargoes to different compartments.