Novel Technologies for intracellular manipulation

 描述（由申请人提供）：该SBIR项目旨在开发用于研究，诊断和最终治疗目的的活真核细胞内部操作和标记的新技术。细胞穿透肽提供了通过真核细胞的质膜递送广泛的用户定义的蛋白质货物的诱人的前景。经由CPP的货物蛋白递送在可递送的生物分子的种类、蛋白表达/递送的快速性和随时间推移的蛋白水平的控制方面具有优于转染的许多优点。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。我们的创新核心技术是基于细胞穿透肽偶联衔接蛋白，允许自发装载几乎任何所需的货物蛋白，快速输送到细胞。我们的CPP适配器技术不仅允许交付更多种类的货物，而且与现有技术相比，在速度和易用性以及安全性方面具有优势。此外，货物从细胞质和系统中的衔接子释放，并且可以作为模块化试剂盒生产，允许使用CPP-衔接子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的特性（例如核，线粒体或溶酶体定位）进行亚细胞递送，并能够同时将多种货物递送到不同的隔室。

 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.