DNA引导基因组使用Natronobacterium gregoryi Argonaute编辑.pptVIP

NgAgo与SpCas9的区别 virtually all organisms / prokaryotes ; bind 5′-phosphorylated ssDNAs and cleave target DNAs / guide RNAs must have a 3′ RNA-RNA hybridization structure for correct Cas9 binding ; no specific sequence on targets required / cleave a target upstream of a PAM. 技术成果 NgAgo can be programmed with ssDNA guides, a precise and efficient tool for genome editing in mammalian cells. 2 Results 2.1 Argonaute derived from N. gregoryi uses DNA guides and cleaves DNA 工作基础：TtAgo/PfAgo在5端磷酸化的单链DNA存在下能够在体外切割DNA靶序列（局限：需要非常高的反应温度，65°C）[1,2]。 Han的灵感：PSI-BLAST，以TtAgo/PfAgo氨基酸序列为对象，在NCBI非冗余蛋白序列数据库