Gene knockin (KI) refers to the process of inserting a segment of DNA sequence at a specific location in the genome, such as adding GFP fluorescence tags to label proteins, or knocking in Cre tool mice in situ, among other applications. For the insertion of small fragments under 1kb, CRISPR/Cas9 has a high efficiency. For the insertion of larger gene fragments, it is necessary to use ES or EPS preparation strategies to ensure project timelines and efficiency.

Figure 1. Knocking in the green fluorescent protein at the C-terminus of the target gene for fusion expression and tracking of the target gene.

1. Technical Process:

Targeting vector design → Vector construction → Microinjection (EPS targeting) → Mouse identification

2. Technical Advantages:

(1) CRISPR is used for the knockin of small fragments.

(2) ES/EPS is used for the knockin of large gene fragments.

(3) Adding various tags to genes, in situ knockin for the preparation of Cre tool mice.