Datenschutz

Transgenic Xenopus strains


DNA CONSTRUCTS
The most important DNA constructs made in the lab are available from addgene:
http://www.addgene.org/Gerhart_Ryffel


CELL LINES
INS-1 and HEK293 cell lines for conditional expression of transgenes as published (Thomas et al 2004, Lucas et al 2005, Senkel et al 2005, Erdmann et al 2007, Senkel et al 2009, Waldner et al 2011, submitted) are available from Sabine Senkel:
sabine.senkel@uni-due.de


TRANSGENIC STRAINS OF XENOPUS LAEVIS
Deposited at the European Xenopus Resource Centre University of Portsmouth
http://www.port.ac.uk/research/exrc
and at the Marine Biological Laboratory (MBL), Woods Hole
http:/www.mbl.edu/xenopus/laevis-stocks


 
(a) REPORTER STRAINS
 

C5: Contains the Cre/FLP reporter LCMV:ECFP(loxP)(FRT)EYFP.
The larvae show blue fluorescence that turns into yellow fluorescence upon Cre or FLP recombinase action (Ryffel et al., 2003;Waldner et al., 2006).

Y2: Contains the Cre reporter CMV:ECFP(loxP)EYFP.
The larvae show blue fluorescence that turns into yellow fluorescence upon Cre recombinase action (Ryffel et al., 2003).

GpL8: Contains the Cre reporter CMV:ECFP(loxP)LacZ.
The larvae show green fluorescence that upon Cre recombinase action turns into ß-galactosidase activity that can be visualized by LacZ staining (Waldner et al., 2006).

 

Tom3: This strain contains a CMV driven red fluorescent protein (tdTomato, kindly provided by Shaner et al., 2004). Adult animals of this strain show ubiquitous and homogenous expression of the red fluorescent protein (Fig. 1) indicating that the CMV promoter can be used to drive transgene expression also at later stages of development (Waldner et al., 2006).

The Figure shows an adult frog of the CMV:tdTomato strain tom3.
A: Head illuminated for red fluorescence.
B and C: Dissected mesonephros of a tom3 animal (right side) or of a control (left side) seen with normal light (B) or red fluorescence (C).

 

(b) EFFECTOR STRAINS
 
328del-4:
Contains the Cre/FLP effector CMV:ECFP(loxP)(FRT)HNF1BP328L329del.
The larvae show blue fluorescence and express upon Cre recombinase action the human mutated HNF1B P328L329 (Roose et al., 2009).

328del-3:
Contains the Cre/FLP effector CMV:ECFP(loxP)(FRT)HNF1BP328L329del and is an independent line from 328del-4 with most similar properties.
The larvae show blue fluorescence and express upon Cre recombinase action the human mutated HNF1B P328L329 (Roose et al., 2009).

A263ins-6:
Contains the Cre/FLP effector CMV:ECFP(loxP)(FRT)HNF1BA263insGG. The larvae show blue fluorescence and express upon Cre recombinase action the human mutated HNF1B A263insGG (Sauert et al., 2011)
 
 
(c) RECOMBINASE STRAINS
 
A7: Contains the CAR-Cre construct that encodes the Cre recombinase coding sequence under control of the cardiac actin promoter. Cre activity has been proven by crossing with the reporter strains C5, Y2 and GpL8 (Waldner et al., 2006).


HSPCre1: This strain contains a Cre recombinase under the heat-shock promoter HSP70. F1 animals of this strain crossed to the C5 reporter strain reveal ubiquitous induction of the EYFP reporter upon heat-shock. DNA recombination was observed as early as one hour after the beginning of heat-shock treatment (Roose et al., 2009).


HSPCre1;C5: These frogs are from a crossing between HSPCre1 and C5. They were heat-shocked in embryogenesis and the yellow larvae (activated EYFP) were grown up.


HSPCre13: This strain contains a Cre recombinase under the heat-shock promoter HSP70. As this strain contains a linked CMV driven gene encoding the red fluorescent protein tdTomato, offspring with Cre recombinase can be easily identified. F1 animals of this strain crossed to the C5 reporter strain reveal ubiquitous induction of the EYFP reporter upon heat-shock (Roose et al., 2009; Sauert et al., 2011).


Reference List
Roose,M., Sauert,K., Turan,G., Solomentsew,N., Werdien,D., Pramanik,K., Senkel,S., Ryffel,G.U., and Waldner,C. (2009). Heat-shock inducible Cre strains to study organogenesis in transgenic Xenopus laevis. Transgenic Res. 18, 595-605.

Ryffel,G.U., Werdien,D., Turan,G., Gerhards,A., Goosses,S., and Senkel,S. (2003). Tagging muscle cell lineages in development and tail regeneration using Cre recombinase in transgenic Xenopus. Nucleic Acids Res. 31, e44.

Sauert,K., Kahnert,S., Roose,M., Gull,M., Brändli,A.W., Ryffel,G.U., and Waldner,C. (2011). Heat-shock mediated overexpression of HNF1b mutations has differential effects on gene expression in the Xenopus pronephric kidney. PLoS. ONE. submitted.

Waldner,C., Roose,M., and Ryffel,G.U. (2009). Red fluorescent Xenopus laevis: a new tool for grafting analysis. BMC. Dev. Biol. 9, 37.

Waldner,C., Sakamaki,K., Ueno,N., Turan,G., and Ryffel,G.U. (2006). Transgenic Xenopus laevis strain expressing cre recombinase in muscle cells. Dev. Dyn. 235, 2220-2228.

 

 

Nach oben