Nlgn3^tm4Sud

The following elements were inserted into intron 1: an FRT site, a duplicate of the 3' end of intron 1 and 5' end of the ATG translation start site-containing exon 2, a full-length rat Nlgn3 cDNA (incorporating mutations (L399A, N400A, D402N, E297A, and K306A) that abolish neurexin binding in paralog Nlg1) fused at codon 780 to the monomeric green fluorescent protein mVenus gene, a splice donor site, an F3 FRT site, a neomycin resistance gene cassette, a tandem FRT/F3 FRT site, and a loxP511 site. A second loxP511 site was inserted into intron 3 to allow for conditional cre-mediated deletion of exons 2 and 3. This allele expresses the mutated Nlgn3-mVenus chimeric protein, with the reporter peptide attached to the cytoplasmic side of the Nlgn3 peptide. If this allele is subjected to flp-mediated recombination, two derivative alleles will be produced: a knockin allele without the neo cassette (recombined via the F3 FRT sites), and a conditional knockout (with the neo cassette and knockin transcript removed) expressing the wild-type peptide (recombined via the FRT sites). qRT-PCR reveals decreased expression levels of the knockin transcript compared to the wild-type. (J:214636)