Anupama K Puppala, Andrew C Nielsen, Maureen R Regan, Georgina E Mancinelli, Renee F DePooter, Stephen Arnovitz, Caspian Harding, Michaele McGregor, Nikolas G Balanis, Ryan Clarke, Brad J Merrill
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引用次数: 0
Abstract
Although genomes encode instructions for mammalian cell differentiation with rich syntactic relationships, existing methods for genetically programming cells have modest capabilities for stepwise regulation of genes. Here, we developed a sequential genetic system that enables transcriptional activation of endogenous genes in a preprogrammed, stepwise manner. The system relies on the removal of an RNA polymerase III termination signal to induce both the transcriptional activation and the DNA endonuclease activities of a Cas9-VPR protein to effect stepwise progression through cascades of gene activation events. The efficiency of the cascading system enables a new dimension for cellular programming by allowing the manipulation of the sequential order of gene activation for directing the differentiation of human stem cells.
尽管基因组编码的哺乳动物细胞分化指令具有丰富的句法关系，但现有的细胞基因编程方法在逐步调控基因方面能力有限。在这里，我们开发了一种顺序遗传系统，能以预编程的方式逐步激活内源基因的转录。该系统依靠去除 RNA 聚合酶 III 终止信号来诱导 Cas9-VPR 蛋白的转录激活和 DNA 内切酶活性，从而通过级联基因激活事件实现逐步推进。级联系统的高效性为细胞编程提供了一个新的维度，它允许操纵基因激活的顺序来指导人类干细胞的分化。
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