FVB/N-Tg(Myh6-AIP/PLN*)46Jded/J

Stock number: 008716
Research areas: Cardiovascular Research, Cell Biology Research, Developmental Biology Research, Internal/Organ Research, Research Tools

Description

SR-AIP transgenic mice carry the α-MHC-AIP₄-SR transgene, with expression of AIP₄ (a tetramer of the CaMKII autocamtide inhibitory peptide AIP) directed to the sarcoplasmic reticulum (SR) of the heart by the murine alpha-myosin heavy chain (α-MHC or Myh6) promoter and a truncated phospholamban (PLB) transmembrane domain (harboring two loss-of-function mutations to prevent direct SERCA inhibition by the fusion protein). The SR-targeted AIP concatemer peptide binds CaMKII, preventing phosphorylation of PLB and subsequent activation of SERCA, thereby increasing diastolic intracellular calcium. These SR-AIP transgenic mice may be useful in studying sarcoplasmic reticulum-targeted CaMKII inhibition, cardiac disease, and the effects of pregnancy upon heart function.

Detailed description

SR-AIP transgenic mice carry the α-MHC-AIP₄-SR transgene. Hemizygous SR-AIP mice are viable and fertile, with expression of AIP₄ (a tetramer of the CaMKII autocamtide inhibitory peptide AIP) directed to the sarcoplasmic reticulum (SR) of the heart by the murine alpha-myosin heavy chain (α-MHC or Myh6) promoter and a truncated phospholamban (PLB) transmembrane domain (harboring two loss-of-function mutations to prevent direct SERCA inhibition by the fusion protein). FLAG epitope expression may be used to identify presence of the fusion protein. The SR-targeted AIP concatemer peptide binds CaMKII, preventing phosphorylation of PLB and subsequent activation of SERCA, thereby increasing diastolic intracellular calcium. Whole heart function and diastolic relaxation are slightly impaired, and pregnancy leads to earlier onset of cardiac hypertrophy. These SR-AIP transgenic mice may be useful in studying sarcoplasmic reticulum-targeted CaMKII inhibition and cardiac disease, as well as the effects of pregnancy upon heart function.

Development

The α-MHC-AIP₄-SR transgene was designed with the 5.5-kb murine alpha-myosin heavy chain (α-MHC or Myh6) promoter, an AIP₄ CaMKII inhibitor domain (encoding a tetramer of the CaMKII autocamtide inhibitory peptide AIP (KKALRRQEAVDAL; a 13-amino acid sequence known to be a highly specific and potent inhibitor of CaMKII)), a FLAG epitope (DYKDDDDK), and a synthetic truncated phospholamban (PLB) transmembrane domain (amino acids 23-52 with L31A/N34A mutations to ensure AIP₄ is targeted to the sarcoplasmic reticulum (SR) without the usual inhibition of the PLB transmembrane domain on SERCA activity). This transgene was injected into pronuclei of FVB/N fertilized mouse oocytes, and transgenic offspring were bred to FVB/N to establish founder line 46 (found to carry an estimated 2 copies of the transgene). These SR-AIP transgenic mice were then bred together for approximately 18-20 generations prior to arrival at The Jackson Laboratory. Upon arrival, SR-AIP mice were bred with FVB/NJ inbred mice (Stock No. 001800) to establish the colony.

Allele

Symbol: Tg(Myh6-AIP/PLN*)46Jded
Name: transgene insertion 46, John R Dedman
MGI accession ID: MGI:3836606
Generation method: Transgenic
Attributes: Inserted expressed sequence
Marker symbol: Tg(Myh6-AIP/PLN*)46Jded
Marker name: transgene insertion 46, John R Dedman
Chromosome: UN
Strain of origin: FVB/N
Expressed genes: Camk2a,calcium/calmodulin-dependent protein kinase II alpha,mouse, laboratory; Pln,phospholamban,mouse, laboratory
Molecular note: The alpha-MHC-AIP4-SR transgene was designed with the 5.5-kb murine alpha-myosin heavy chain (alpha-MHC or Myh6) promoter, an AIP4 CaMKII inhibitor domain (encoding a tetramer of the CaMKII autocamtide inhibitory peptide AIP (KKALRRQEAVDAL; a 13-amino acid sequence known to be a highly specific and potent inhibitor of CaMKII)), a FLAG epitope (DYKDDDDK), and a synthetic truncated phospholamban (PLB) transmembrane domain (amino acids 23-52 with L31A/N34A mutations to ensure AIP4 is targeted to the sarcoplasmic reticulum (SR) without the usual inhibition of the PLB transmembrane domain on SERCA activity). This transgene was injected into pronuclei of FVB/N fertilized mouse oocytes, and transgenic offspring were bred to FVB/N to establish founder line 46 (found to carry an estimated 2 copies of the transgene).