Easi-CRISPR: Generating Knock-In and Conditional Mouse Models
Type
Blog Post
...of conventional gene targeting is lowered to about 2-3 months. But certain models remain more difficult...included in the final ssDNA template.
Figure 2: Overview of ivTRT to generate an ssDNA repair template... floxed conditional allele requires you to target 2 loxP sites to intronic regions 0.5 - 0.8 kb apart....Typically, Easi-CRISPR works best with insertions of 2 kb or less. Luckily, most of the commonly used cassettes... induction systems (ttA or rtTA) are between 1 to 2 kb in length. There are plenty of Addgene plasmid ...create the ssDNA construct as described in Figure 2.
You can also use Addgene plasmids to purify Cas9...desired mutation in mouse ES cells, select for the mutation and inject the ES cells into a mouse embryo to ...