B26

B26: cdh5-optoα1AR-IRES-lacZ Strain Information Sheet

 

Strain Name:

Common Name: cdh5-optoα1AR-IRES-lacZ

CHROMusTM designation and lines available: R24:B26

Jackson Stock number:  Jax# 033343

Development: The rhodopsin based effector, optoα1AR which on activation generates the second messenger diacylglycerol/IP3, was inserted at the ATG site of the cdh5 gene in the BAC RP23-453P1 (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 # expressors
64 9 tbd 1+

Description: The rhodopsin based effector, optoα1AR is expressed under control of the cdh5 promoter, directing expression to endothelial cells. Expression is observed in endothelial cells of vessels. An internal ribosome entry site (IRES) allows for co-expression of lacZ as a visual indicator of expression patterns. This mouse is useful for activating second messengers in endothelial cells.

Phenotypic Data:

Functional analysis is ongoing.  LacZ staining of heart, brain and lung shows appropriate expression patterns.

cdh5-optoa1AR-IRES-lacZ staining of heart and lung

cdh5-optoa1AR-IRES-lacZ staining of heart and lung

Genotyping Protocol:

Primer B101 GGGTTGGTCCCGCTATATTC
Primer B102 GAAGGCAGGGATGGTCATAAA
Expected size of product: 478bp
Cycling conditions:

  1. 94oC 3 minutes
  2. Repeat 30 times
    1. 30 sec @ 94oC
    2. 30 sec @ 58oC
    3. 30 sec @ 72oC
  3. 72oC 5 minutes
  4. 10oC hold

PCR results:

PCR results for optoa1AR B101 x B102

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 cdh5-optoα1AR-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 cdh5-optoα1AR-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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