B17

HCN4-CatCh-IRES-lacZ Strain Information Sheet

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Strain Name:

Common Name: HCN4-CatCh-IRES-lacZ

CHROMusTM designation and lines available: R24:B17

Jackson Stock number: NA

Development: The rhodopsin based effector, CatCh was inserted at the ATG site of the HCN4 gene in the BAC RP23-281H22 (CHORI) through homologous recombination. The resulting recombinant BAC was injected into the male pronucleus of fertilized oocytes which were then implanted into pseudo pregnant females. Resulting offspring were screened for the presence of the transgene (founders). Colonies were established from each founder and tested for expression.

Transgenic Numbers:

# pups born # founders # expressers
17 2 2

Description: The rhodopsin based effector, CatCh is expressed under control of the HCN4 promoter, directing expression to the SA node and the AV node of the heart and the nervous system (brain and spinal cord). Expression is observed in the developing 13.5 dpc heart.   An internal ribosome entry site (IRES) allows for co-expression of lacZ as a visual indicator of expression patterns.

Phenotypic Data:

Line 1 13.5dpc embryo open chest showing lacZ staining of sinus venosus.

Line 1 13.5dpc embryo open chest showing lacZ staining of sinus venosus.

Line 1 13.5dpc embryo showing lacZ staining in the sinus venosus.

Line 1 13.5dpc embryo showing lacZ staining in the sinus venosus.

Line 2 13.5 dpc embryos stained for lacZ and dissected to show the open chest with the sinus venosus and left ventricle staining blue in transgenic positive animals.

Line 2 13.5 dpc embryos stained for lacZ and dissected to show the open chest with the sinus venosus and left ventricle staining blue in transgenic positive animals.

Line 1, 10.5 dpc embryos lacZ staining

Line 1, 10.5 dpc embryos lacZ staining


 

Additional data detailing CatCh activity will be added.

Genotyping Protocol:

Primer B107 GGA GAT CTA TGT GTG CGC TAT C
Primer B108 CCA GCA GTT CTT CGA CAT CA
Expected size of product: 535bp

Cycling conditions:

94oC 3 minutes
Repeat 30 times
30 sec @ 94oC
30 sec @ 58oC
30 sec @ 72oC
72oC 5 minutes
10oC hold

Terms of Use: Please contact chromus@cornell.edu for information on MTA and fees.

Acknowledgement: Please inform us of any publications resulting from the use of this mouse and use the following statement to acknowledge their development:

“The mouse strain HCN4-CatCh-IRES-lacZ was developed by CHROMusTM; funded by NIH R24HL120847.”

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