|Characteristics||CRISPR/Cas9 generated medaka mutant carrying a deletion in osterix/Sp7 (transcript variant 1) in a osx:mCherry transgenic background, with premature and mature osteoblasts expressing mCherry. Heterozygous mutant larvae showed slightly reduced numbers of osteoblasts in shortened neural arches with asymmetrical length. However, this arch phenotype is sometimes too sultle to observe. Thus, mutant genotype of an adult fish needs to be confirmed by RFLP. Homozygous mutant larvae showed strong intramembranous bone defects in cranial structures, arches as well as caudal fin rays. Absence of osx:mCherry positive cells can be observed in structures with bone defects. Homozygous mutant larvae have high lethal rate and are not viable.|
|Special affairs [Address]||
■ The RECIPIENT shall refer the following publication in any PUBLICATIONS.
(Name of the Publication: Yu T, Graf M, Renn J, Schartl M, Larionova D, Huysseune A, Witten PE, Winkler C. (2017). A vertebrate-specific and essential role for osterix in osteogenesis revealed by gene knockout in the teleost medaka. Development 144, 265-271.)
■ 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|
|Frozen sperm in NIBB||○|
|Deposited by||National University of Singapore|
osx_mCherry transgene map
A 4.1-kb fragment (including a putative Runx2 binding site) upstream of the osterix (osx) translational start site from genomic DNA was cloned in front of mCherry into a vector with flanking I-SceI meganuclease restriction sites (for details see Renn and Winkler, Dev Dyn 2009).
TG1138_Dorsal view of embryo
(A') osx:mcherry-expressing osteoblasts in wildtype sibling. Dorsal view of embryo at 12 dpf. (B') osx:mcherry-expressing osteoblasts in osx-/- mutant, dorsal view at 15 dpf. Note reduction of osx-positive osteoblasts skull bones such as branchiostegal rays and parasphenoid. (A') and (B') are merged images from brightfield and fluorescent channels. Boxes indicate areas with auto-fluorescent pigment cells.
TG1138_Lateral view of embryo
(C) Lateral view of wildtype embryo at 12 dpf with osx:mCherry-expressing osteoblasts. (D) Lateral view of osx-/- mutant with reduced numbers of osx:mCherry expressing cells. Note that image (D) was taken with extended exposure time to achieve sufficient signal. (C') and (D') show higher magnification of vertebral body region boxed in (C) and (D).
|Document (PDF)||Genotyping protocol_TG1138_osx_tv1 mutant|