CRISPR Knock in是什么?
CRISPR/Cas9敲入
a precise form of repair (homology-directed repair, HDR) by providing a DNA sequence that the cell can use as a repair template to insert (knock in) a matching DNA sequence into the break.
Example application include modification of a promoter sequence or gene, insertion of a exogenous reporter (e.g., a fluorescent protein), or creation of a clinically relevent SNP for a disease model.
Knockin at one copy fo the gene / sequence is usually accompanied by a knockout / indel at the second copy
Both copies are cut by the Cas9/sgRNA complex but one is likely repaired imprecisely using the NHEJ pathway, creating an indel
CRISPR/Cas9敲入
提供一段DNA序列, 细胞可将其作为修复模板插入匹配的DNA序列进入断裂位置, 是一种精确的修复 (同源定向修复)
应用举例包括: 修饰启动子序列, 基因, 插入外源性的报告子(荧光蛋白), 或为一种疾病模型制造临床相关的单核苷酸多态性
敲入基因/序列的一个拷贝通常伴随第二个拷贝的敲除/插入确实
两个拷贝都被切割, 但其中一个可能通过NHEJ途径不精确的修复, 从而产生了indel (插入/ 删除)
