确保SgRNA能够靶向目标基因的特定位置，并且与Cas9蛋白兼容。SgRNA scaffold设计:sgRNA scaffold是一个特定的...
在张峰实验组sgrnascaffold序列中,除过序列末端有5个连续的t碱基,在sgrnascaffold起始位点即crrna的repeat序列也有4个连续的t碱基,申请人推测该序列会影响转录提前终止,从而降低sgrnascaffold转录,也就间接影响cas9/sgrna复合体的基因编辑活力。张峰团队为了解决该问题而将tttt改为tatt碱基,但是通过t7endonucleaseⅰ酶切鉴定...
As a proof of the concept, we established a chain reaction through the repair of sgRNA scaffold mutations in 293T cells. Importantly, the results obtained in yeast or in vitro were not consistent with those in mammalian cells, suggesting that the in vivo chain reactions need to be optimized ...