Embodiments disclosed herein are directed to engineered CRISPR-Cas effector proteins that comprise at least one modification compared to an unmodified CRISPR-Cas effector protein that enhances binding of the of the CRISPR complex to the binding site and/or alters editing preference as compared to wild type. In certain example embodiments, the CRISPR-Cas effector protein is a Type V effector protein. In certain other example embodiments, the Type V effector protein is Cpf1. Embodiments disclosed herein are directed to viral vectors for delivery of CRISPR-Cas effector proteins, including Cpf1. In certain example embodiments, the vectors are designed so as to allow packaging of the CRISPR-Cas effector protein within a single vector. There is also an increased interest in the design of compact promoters for packing and thus expressing larger transgenes for targeted delivery and tissue-specificity. Thus, in another aspect certain embodiments disclosed herein are directed to delivery vectors, constructs, and methods of delivering larger genes for systemic delivery.
本文公开的实施方案涉及工程化的 CRI
SPR-Cas 效应蛋白,与未修饰的 CRI
SPR-Cas 效应蛋白相比,它包含至少一种修饰,与野生型相比,这种修饰增强了 CRI
SPR 复合物与结合位点的结合和/或改变了编辑偏好。在某些示例性实施方案中,CRI
SPR-Cas效应蛋白是V型效应蛋白。在某些其他示例实施方案中,V 型效应蛋白是 Cpf1。本文公开的实施方案涉及用于递送包括 Cpf1 在内的 CRI
SPR-Cas 效应蛋白的病毒载体。 在某些示例性实施方案中,载体的设计允许在单个载体中包装 CRI
SPR-Cas 效应蛋白。此外,人们对设计紧凑型启动子也越来越感兴趣,这种启动子可用于包装,从而表达更大的转
基因,以实现定向传递和组织特异性。因此,在另一个方面,本文公开的某些实施方案针对的是递送载体、构建体和递送较大
基因用于系统递送的方法。