B6.Cg-Sema6a^tm2Alk/J

Stock number: 039871
Product name: Sema6A^flox
Strain synonyms: Sema6A^HA-F
Research areas: Cancer Research, Developmental Biology Research, Neurobiology Research, Research Tools, Sensorineural Research

Description

The Sema6A^flox strain has a floxed exon 3 in the sema domain, transmembrane domain (TM), and cytoplasmic domain, (semaphorin) 6A (Sema6A) gene. Sema6A is a transmembrane protein that has many implicated functions, including axon guidance, cell migration, cancer progression, and myelination. The line is useful for studies of Sema6A in neural development and axonal guidance, in addition to studies addressing the development and function of multiple organ systems.

Of note, the donating investigator has made available mice with Sema6A^flox [3xHA::loxP-exon3-loxP; Stock No. 039871] and its germline Cre recombined derivative Sema6A^null [retaining 3xHA with exon3 deleted; Stock No. 039872].

Detailed description

The Sema6A^flox conditional knock-out mice possess an N-terminus hemagglutinin (HA) tag and floxed exon 3 in the sema domain, transmembrane domain (TM), and cytoplasmic domain, (semaphorin) 6A (Sema6A) gene. The homozygous mice are viable and fertile before Cre-recombination. The Sema6A^flox conditional allele produces an N-terminal, HA-tagged, fully functional SEMA6A protein (HA-Sema6A) in the absence of Cre recombinase. The donating investigator reported that the HA-Sema6A immunolabeling faithfully and reproducible reports Sema6A expression. Sema6A is expressed in the developing nervous system and other organ systems. It is expressed by selective retinal ganglion cells and is believed to be important in regulating neuron patterning and axon guidance. The line is useful for studies of Sema6A in neural development and axonal guidance, in addition to studies addressing the development and function of multiple organ systems.

Of note, the donating investigator has made available mice with Sema6A^flox [3xHA::loxP-exon3-loxP; Stock No. 039871] and its germline Cre recombined derivative Sema6A^null [retaining 3xHA with exon3 deleted; Stock No. 039872].

Development

A targeting construct was designed to insert a 3x hemagglutinin (HA) tag downstream of the signal peptide in exon 2 at the N-terminus of the extracellular domain of Sema6A and loxP sites in introns flanking exon 3. The targeting construct was injected into (129S6 x C57BL/6J)F1-derived ES cells. The frt-flanked neo cassette was removed by transient FLP recombinase expression in the ES cells. Then, ES clones with correct insertion were verified by sequencing and introduced into recipient mice blastocysts. The chimeric mice were bred with C57BL/6 mice. The resulting mice were then backcrossed more than 8 generations to C57BL/6J mice and maintained in C57BL/6 genetic background by donating laboratory. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes.

Allele

Symbol: Sema6a^tm2Alk
Name: targeted mutation 2, Alex L Kolodkin
MGI accession ID: MGI:7660920
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Epitope tag
Synonyms: HA-Sema6a^fl; Sema6a^HA-F
Marker symbol: Sema6a
Marker name: sema domain, transmembrane domain (TM), and cytoplasmic domain, (semaphorin) 6A
Marker synonyms: A730020P05Rik; HT018; sema; SEMA6A1; Sema6A-1; Semaq; VIa
Chromosome: 18
Strain of origin: (129S6/SvEvTac x C57BL/6J)F1
Molecular note: The targeting construct inserted an hemagglutinin (HA) tag is knocked in downstream of the signal peptide in exon 2 at the N-terminus of the extracellular domain and floxed exon 3. The frt-flanked neo cassette was removed by crossing the FLP deleter line.