Single transgenic expressing the osteoclast-inducing factor Receptor Activator of Nuclear Factor kappa B ligand (RANKL) and CFP under bidirectional control of a heat shock-inducible promoter. Upon Rankl induction by heat-shock (incubating embryos at 39°C for 2 hours) at 9dpf, in rankl:HSE:CFP larvae, a severe loss in mineralized matrix of most embryos overexpressing Rankl resulting in an osteoporotic phenotype can be observed; with losses varying in extent (Fig. 7A-D in To TT et al., 2012).
|Notes||For screening of transgenic carriers, a short early heat shock should be conducted to induce CFP (but avoid any osteoporotic effect). Ideally, a heat shock is done at 2 dpf for 1 hour at 39C. The CFP signal can be observed 2 hours after heat shock for 20 min at 39C. Stronger signal can be seen the next day after. Heat shock after 8 dpf will induce osteoclast formation and osteoporotic phenotype.|
|Special affairs [Address]||
■ The RECIPIENT shall refer the following publication in any PUBLICATIONS.
(Name of the Publication: RENN, Joerg; WINKLER, Christoph. Osterix‐mCherry transgenic medaka for in vivo imaging of bone formation. Developmental Dynamics, 2009, 238.1: 241-248.
TO, Thuy Thanh, et al. Rankl-induced osteoclastogenesis leads to loss of mineralization in a medaka osteoporosis model. Development, 2012, 139.1: 141-150
RENN, Jörg, et al. A col10a1: nlGFP transgenic line displays putative osteoblast precursors at the medaka notochordal sheath prior to mineralization. Developmental biology, 2013, 381.1: 134-143.)
■ The RECIPIENT shall give proper credit to the DEVELOPER in any publications reporting results of research using any derivatives developed from the BIOLOGICAL MATERIALS (hereinafter referred to as “the DERIVATIVES”), and shall procure other institutionsto which the DERIVATIVES are provided (including when provided through the NBRP) to do the same.
|Organization||National Institute for Basic Biology|
|Deposited by||National University of Singapore|
TG1135_RANKL HSE CFP 10dpf
Lateral view of medaka at 10 dpf, 1 day after heatshock (2 hours, 39 C). CFP fluorescence throughout body.
TG1135_RANKL HSE CFP_transgene map
The full-length open reading frame (ORF) of medaka rankl (786 nt, 262 aa) was cloned into BamHI-XhoI sites of pSGH2 (a gift from Thomas Czerny lab, Bajoghli B. et al., Dev Biol. 2004 Jul 15;271,416-430, 2004. pSGH2 contains a bidirectional heat-shock promoter (8x HSE) driving expression of CFP and has two flanking I-SceI sites for transgenesis. For details see To TT et al., 2012. The RANKL:HSE:CFP construct contains a bidirectional heat-shock promoter driving simultaneous expression of rankl and CFP in all cells of the body. The vector backbone sequence is not available.