B6.129X1(Cg)-Bcl2^tm1.1Sjk/J

Stock number: 018430
Research areas: Diabetes and Obesity Research, Metabolism Research

Description

B cell leukemia/lymphoma 2 (BCL2) is an anti-apoptotic, anti-autophagy protein that inhibits autophagy by interacting with Beclin 1 (BECN1). Alanine substitutions in these mice prevent stimulus-induced disruption of the BCL2-BECN1 complex and autophagy activation. These mice may be useful in studies of induced autophagy, metabolism, and exercise.

Detailed description

B cell leukemia/lymphoma 2 (BCL2) is an anti-apoptotic, anti-autophagy protein that inhibits autophagy by interacting with Beclin 1 (BECN1). Disruption of the BCL2-BECN1 complex is necessary for stimulus (exercise or starvation)- induced autophagy. Bcl2^AAA mice carry alanine substitutions in three conserved phosphorylation sites (T69A, S70A, S84A) that prevent stimulus-induced disruption of the BCL2-BECN1 complex and autophagy activation. Homozygous mice are viable and fertile. During acute exercise, mice exhibit decreased endurance, impaired insulin sensitivity, altered glucose metabolism and impaired protection against glucose intolerance from high fat diet. Bcl2^AAA mice may be useful in studies of induced autophagy, metabolism, and exercise.

Development

A targeting vector containing an frt-flanked puromycin resistance cassette was used to insert alanine substitutions into amino acid residues 69 (threonine), 70 (serine), and 84 (serine) of the Bcl2 gene (Bcl2^AAA). The alanine substitutions were directed to three conserved phosphorylation sites of the exon 2 loop region. The construct was electroporated into 129X1/SvJ-derived RW4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and resulting chimeric mice were bred with B6;SJL-Tg(ACTFLPe)9205Dym/J to remove the puro cassette. Mice were backcrossed to C57BL/6J for more than 10 generations.

Allele

Symbol: Bcl2^tm1.1Sjk
Name: targeted mutation 1.1, Stanley Korsmeyer
MGI accession ID: MGI:5313428
Generation method: Targeted
Marker symbol: Bcl2
Marker name: B cell leukemia/lymphoma 2
Chromosome: 1
Strain of origin: 129X1/SvJ
Molecular note: Nucleotide substitutions in exon 2 were used to replace the threonine at 69 and serines at positions 70 and 84 with alanines (T69A, S70A and S84A). Flp-mediated recombination removes the puro cassette inserted downstream of the modified exon 2.