STOCK Hyal3^tm1Btr/J

Stock number: 040597
Product name: Hyal3 KO
Research areas: Cell Biology Research, Developmental Biology Research, Research Tools

Description

Hyal3^-/- knock-out mice carry a neomycin resistance cassette (neo) inserted into exon 2 of the (Hyal3) gene, inactivating full-length transcription. These mice may be useful for studying the role of Hyal3 in breakdown of hyaluronan (HA), an abundant component of the vertebrate extracellular matrix.

Detailed description

The Hyal3^-/- knock-out mice carry a neomycin resistance cassette (neo) inserted into exon 2 of the (Hyal3) gene, inactivating full-length transcription.

Hyaluronidases are endoglycosidases that initiate the breakdown of hyaluronan (HA), an abundant component of the vertebrate extracellular matrix. In mice, there are 7 hyaluronidases paralogs (6 in human) clusters on Chromosome 6 and Chromosome 9 suggesting redundant/overlapping function with other HYAL genes.

The Hyal3 knockout (Hyal3^-/-) mice are viable, fertile, and exhibit no gross phenotypic changes. RT-PCR analysis showed that no wildtype HYAL3 transcript is expressed from this allele, and that overlapping Naa80 transcripts are not altered in the testis, but the levels are reduced by about 20% in the liver. X-ray analysis, histological studies of joints, whole-body weights, organ weights and the serum HA levels of Hyal3^-/- mice were normal. No glycosaminoglycan (GAG) accumulation was detectable, suggesting that under normal circumstances, HYAL3 does not contribute significantly to HA degradation, or its functional redundancy exists among other HYAL genes. These mice may be useful for studying the role of Hyal3 in extracellular matrix hyaluronan (HA) remodeling during development and/or cancer progression.

Development

A neomycin resistance cassette (neo) was inserted into exon 2 of the Hyal3 gene at the NsiI site 40 bp downstream of its initiation codon (Reference sequence: NM_178020.3). This targeting construct was designed to disrupt Hyal3 while having minimal impact on an overlapping gene, Naa80 (N(alpha)-acetyltransferase 80, NatH catalytic subunit; human synonym: FUS2) which second exon is embedded within the first intron of the Hyal3 gene. The targeting vector was linearized and introduced into (129X1/SvJ x 129S1/Sv)F1-Kitl⁺ derived R1 embryonic stem (ES) cells. Correctly targeted ES cell clones were identified by Southern hybridization and aggregated with CD1 morulae. Male chimeras transmitting the targeted Hyal3 allele were identified for two of the ES cell clones and were used to establish two independent lines of mixed background with CD1 mice. One of the male chimeras transmitting the Hyal3 targeted allele in their germline was also crossed with 129S1 (Charles River Laboratories, Quebec, Canada) female mice to generate CD1/C129S background. The other line was maintained as an outcross CD1/C129S background. Then later, the line was crossed onto a C57BL/6N background to generate 129/C57BL/6N heterozygous mice. These mice were maintained by heterozygote intercrosses until the donating laboratory froze sperm. Upon arrival at The Jackson Laboratory, to establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ (Stock No. 005304) oocytes.

Allele

Symbol: Hyal3^tm1Btr
Name: targeted mutation 1, Barbara Triggs-Raine
MGI accession ID: MGI:6108181
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Hyal3
Marker name: hyaluronoglucosaminidase 3
Marker synonyms: HYAL-3; LUCA3; LUCA-3
Chromosome: 9
Strain of origin: (129X1/SvJ x 129S1/Sv)F1-Kitl⁺
Molecular note: A neomycin resistance cassette (neo) was inserted into exon 2 of the gene at the NsiI site. This targeting construct was designed to disrupt Hyal3 while having minimal impact on an overlapping gene, Nat6 (N-acetyltransferase 6), which second exon is embedded within the first intron of this gene. RT-PCR analysis showed that no wild type transcript is expressed from this allele, and that Fus2 trasncripts are not altered in testis, but the levels are reduced about 20% in liver.