TARGATT / Hipp11
Please see the relevant primary literature below from the Luo Lab at Stanford regarding the identification of the transcriptionally inactive Hipp11 locus and the generation of attP RMCE mice.
Essentially, an attB flanked (or single attB-site containing) plasmid, in the presence of Phi31 integrase protein, will recombine through RCME at the modified Hipp11 locus to yield a single copy, heterozygous, transgenic mouse.
Applied Stem Cell sells the attB flanked and single attB-site containing plasmids. Of consideration is how you will remove the backbone of your plasmid (if applicable). The attP engineered mouse line (available from Charles River) has a FRT5 site downstream of the endogenous attP site (hint). The kit also comes with Phi31o integrase mRNA.
The Phi31o cDNA is also available from addgene, as are several vectors that are excellent for making stable, capped mRNA (such as pCS2).
Alternatively, one could target the Hipp11 locus by CRISPR to eliminate the need for the integrase or the attP-mice.
We've modified the original genotyping protocol to allow for PCR-based screening of founder lines and offspring.
Relevant Literature
Tasic et al., 2011
Hippenmeyer et al., 2010
