Review Article

New Turns for High Efficiency Knock-In of Large DNA in Human Pluripotent Stem Cells

Figure 1

Schematic overview of HDR-, NHEJ-, MMEJ-, and HMEJ-based knock-in. (a) HDR-based knock-in requires long homology arms. (b) NHEJ-based knock-in, which requires the linearization of donor template rather than the homology arms. (c) MMEJ-based knock-in requires short homology arms (usually less than 50 bp). NHEJ-mediated knock-in might happen due to the presence of linearized donor template. (d) HMEJ-based knock-in requires the linearized donor template with long flanking arms. This method may trigger HDR-, SSA-, and NHEJ-based knock-in and yield combinatory outcome.