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 10.5 and 13.5 dpc heart, neonates and adult mice.   An internal ribosome entry site (IRES) allows for co-expression of lacZ as a visual indicator of expression patterns.

Phenotypic Data:

The heart rate of the HCN4-CatCh-IRES-lacZ mouse can paced by laser stimulation of the SA node in the adult (data not shown).  Additionally, lacZ staining shows the extent of the expression of the transgene in all stages of development.

LacZ staining of the heart of PN4 neonates, followed by tissue clearing, shows expression within the intricate network of HCN4 expressing, specialized conducting cells lining both the left and right ventricles as well as the SA and AV nodes.

LacZ staining of the heart of PN4 neonates, followed by tissue clearing, shows expression within the intricate network of HCN4 expressing, specialized conducting cells lining both the left and right ventricles as well as the SA and AV nodes.

HCN4-CatCh-IRES-lacZ adult mouse showing lacZ staining of sections in the heart

HCN4-CatCh-IRES-lacZ adult mouse showing lacZ staining of sections in the heart A) right atrium B) AV junction C) lining of the left ventricle

 

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 1, 10.5 dpc embryos lacZ staining

Line 1, 10.5 dpc embryos lacZ staining

 

 

 

 

 

 

 

 

 

 

 

 

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.

So as to appropriately acknowledge and credit the scientists who have produced the HCN4-CatCh-IRES-lacZ mouse strain, it is expected that you will include these individuals in any publication that is submitted prior to the initial publication of this line from the Kotlikoff laboratory.

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 which is supported by the National Heart Lung Blood Institute of the National Institute of Health under award number R24HL120847.

 

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