Mice carrying the Gbatm1.1Mjff allele were bred to mice carrying the Tg(Thy1-SNCA)15Mjff transgene to generate this double mutant line.
For the Gbatm1.1Mjff allele, a targeting vector containing a FRT site flanked NEO cassette, and a F3 FRT site flanked PurO cassette was used to introduce the D427V point mutation into exon 10. The FRT site flanked NEO cassette was inserted into intron 5 and the F3 FRT site flanked PurO cassette was inserted into intron 8. In addition, loxP sites were inserted to flank exons 6 through 8. The construct was electroporated into C57BL/6NTac derived Art B6 3.6 embryonic stem (ES) cells. Correctly targeted ES cells were injected into BALB/c blastocysts. The resulting chimeric animals were crossed to FLP recombinase expressing females on the C57BL/6NTac background, C57BL/6NTac-Tg(CAG-Flpe)2Art, to remove the selection cassettes. These mice no longer carry the FLP recombinase allele. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6NJ (Stock No. 005304) at least once to establish the colony.
The mThy1-hSNCA transgene consists of the human a-synuclein (SNCA) gene driven by a mouse thymus cell antigen 1 (Thy-1) promoter. The construct was microinjected into fertilized C57BL/6NTac oocytes and males from founder line 15 were bred to C57BL/6NTac females to establish the colony. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6NJ inbred mice (Stock No. 005304) for at least one generation.
