Shh Cre ; Pparg Knockout Mice
- These mice lack superficial layer
B6.Cg-Lamc2 tm1Uit /DcrJ
- These mice lack expression of the Lamc2 gene, which encodes the β subunit of laminin 5 – a matrix glycoprotein found in basement membranes. A complete description is available from Jackson Laboratories.
Lama3 conditional knockout mouse
- These mice expressed a floxed Lam3 gene on exon 42. Lam3 encodes the α3 subunit of laminin (Urich et al., J Cell Sci 2011).
Cell Specific Gene Expression In Vivo
- Uracil phosphoribosyltransferase (UPRT) was inserted into a Rosa-26 targeting vector pBigT-DEST preceded by a floxed-stop motif and followed by a transcriptional stop cassette “PGK-neo-pA-3xpA”, preventing downstream expression.
- Because mammalian cells do not have functional UPRT, the construct directs cell-specific incorporation of 4-ThioUracil during mRNA synthesis.
- The labeled mRNA can then be biotinylated in vitro and isolated to allow cell-specific gene expression analysis. UPRT construct from Drs. Michael Cleary and Christopher Doe.
- Mouse Lrp2 is a huge gene that is over 200 kbp.
- Using recombineering, we replaced the translational STOP (TAG) with “P2a-CreERT2-frt-neo-frt” cassette. The entire “P2a-CreERT2-frt-neo-frt” cassette as well as the 5’ and 3’ of the insertion site were completely sequenced to verify the sequencing read.
- the resulting MC mice are maintained as homozygote. “P2a” allows expression of Lrp2 and CreERT2 from the modified allele.
- The M5C mouse was made by replacing the translational start (ATG) of Lrp2 with the“CreERT2-frt-neo-frt” cassette.
- After removing the Neo cassette, the M5C mice are maintained as heterozygote.
- Expression of CreERT2 in this case will inactivate Lrp2.
- TH-CreERT was also generated by inserting P2a-CreERT2-frt-neo-frt” cassette in the 3’UTR to generate a functional TH and a functional CreERT2.
For more information on these models, please contact us.