Targeting the RORγ locus. (A) Schematic
representation of the mouse RORγ gene locus, the pPNT-ΔRORγ
targeting vector, and the recombination at the RORγ locus. In the
targeted locus the region from exon 3 through exon 6 is deleted. DBD,
DNA-binding domain; LBD, ligand-binding domain. (B)
Diagnostic Southern blot analysis. Genomic DNA was cut by
EcoRV, electrophoresed, and hybridized to the
3′-flanking probe E (indicated in A), which detected
fragments of the expected size of 4.0 kb for wild-type and 5.0 kb for
the mutant allele. Lanes indicate digested DNA from wild-type (wt;
+/+), heterozygous (+/−), and homozygous mutant (−/−) mice.
(C and D) Northern blot analysis was
performed with RNA isolated from liver (L), kidney (K), and thymus of
wt, RORγ+/−, and RORγ−/− mice by using
either a probe encompassing the deleted region encoded by exons 3–6
(C) or a full-length RORγ probe (D).