Basic Information

Strain Name
C57BL/6-Camk2atm1(icre,EGFP)Bcgen/Bcgen
Stock Number
110780
Common Name
B-Camk2a-iCre-EGFP mice
Gene symbol and name
Camk2a, calcium/calmodulin-dependent protein kinase II alpha
Background
C57BL/6
Coat color
Black
Application
• Function research of genes • Function research of synaptic plasticity and Long Term Potentiation (LTP) in hippocampal networks
NCBI Gene ID
Development
A targeting vector was designed to insert an P2A-iCre-P2A-EGFP cassette before the calcium/calmodulin-dependent protein kinase II alpha (Camk2a) stop codon. A Frt-flanked neomycin resistance (Neo) gene was inserted into the intron16-17 of Camk2a gene. The targeting vector was electroporated into C57BL/6-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were bred to FLP transgene mice to delete the neomycin selection cassette. FLP transgene was removed by crossing with wild-type C57BL/6 mice. These mice were maintained on a C57BL/6 background.

Description

Mice homozygous for the Camk2a-iCre are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. In this strain, iCre recombinase expression is under the control of Camk2a promoter. When crossed with a strain containing a loxP-site flanked sequence of interest, Cre-mediated recombination will
result in deletion of the floxed sequence in the offspring.

Gene editing strategy

A targeting vector was designed to insert an P2A-iCre-P2A-EGFP cassette before the calcium/calmodulin-dependent protein kinase II alpha (Camk2a) stop codon. A Frt-flanked neomycin resistance (Neo) gene was inserted into the intron16-17 of Camk2a gene. The targeting vector was electroporated into C57BL/6-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were bred to FLP transgene mice to delete the neomycin selection cassette. FLP transgene was removed by crossing with wild-type C57BL/6 mice. These mice were maintained on a C57BL/6 background.

Phenotype analysis

The flow results showed that the mouse brain could express ICRE and function normally.

The single positive cells of TDTomato were found in testis, heart, liver, spleen, lung and kidney by flow cytometry.The experiment results with Jackson Lab B6. Cg – Tg (Camk2a – cre) STL T29-1 / J (Stock No: 005359 | T29-1) multiple organ test in mice to express the result of the agreement.The IF results of the fluorescent microscopic imaging of the mouse brain showed that the red fluorescent protein of TDTomato was successfully expressed in the mouse brain, and the expression was significantly increased in some areas, especially in the structure of the dentate gyrus, which was highly expressed.

References

  1. Mayford M, Bach ME, Huang YY, Wang L, Hawkins RD, Kandel ER. Control of memory formation through regulated expression of a CaMKII transgene. Science. 1996 Dec 6;274(5293):1678-83. [PMID: 8939850]
  2. Kellendonk C, Tronche F, Casanova E, Anlag K, Opherk C, Schütz G. Inducible site-specific recombination in the brain. J Mol Biol. 1999 Jan 8;285(1):175-82. [PMID: 9878397]
  3. Mantamadiotis T, Lemberger T, Bleckmann SC, Kern H, Kretz O, Martin Villalba A, Tronche F, Kellendonk C, Gau D, Kapfhammer J, Otto C, Schmid W, Schütz G. Disruption of CREB function in brain leads to neurodegeneration. Nat Genet. 2002 May;31(1):47-54. [PMID: 11967539]
  4. Minichiello L, Korte M, Wolfer D, Kühn R, Unsicker K, Cestari V, Rossi-Arnaud C, Lipp HP, Bonhoeffer T, Klein R. Essential role for TrkB receptors in hippocampus-mediated learning. Neuron. 1999 Oct;24(2):401-14. [PMID: 10571233]
  5. Saito H, Yoshida T, Yamazaki H, Suzuki N. Conditional N-rasG12V expression promotes manifestations of neurofibromatosis in a mouse model. Oncogene. 2007 Jul 12;26(32):4714-9. [PMID: 17237809]
  6. Küry S, van Woerden GM, Besnard T. De Novo Mutations in Protein Kinase Genes CAMK2A and CAMK2B Cause Intellectual Disability. Am J Hum Genet. 2017 Nov 2;101(5):768-788. [PMCID: PMC5673671]
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