A systematic genome-wide screen for mutations affecting organogenesis in Medaka, Oryzias latipes

Furutani-Seiki, Makoto; Sasado, Takao; Morinaga, Chikako; Suwa, Hiroshi; Niwa, Kenta; Yoda, Hiroki; Deguchi, Tomonori; Hirose, Yasuyuki; Yasuoka, Akihito; Henrich, Thorsten; Watanabe, Tomomi; Iwanami, Norimasa; Kitagawa, Daiju; Saito, Kota; Asaka, Satoshi; Osakada, Masakazu; Kunimatsu, Sanae; Momoi, Akihiro; Elmasri, Harun; Winkler, Christoph; Ramialison, Mirana; Loosli, Felix; Quiring, Rebecca; Carl, Matthias; Grabher, Clemens; Winkler, Sylke; Bene, Filippo Del; Shinomiya, Ai; Kota, Yasuko; Yamanaka, Toshiyuki; Okamoto, Yasuko; Takahashi, Katsuhito; Todo, Tomoki; Abe, Keiko; Takahama, Yousuke; Tanaka, Minoru; Mikami, Hiroshi; Katada, Toshiaki; Nishina, Hiroshi; Nakajima, Noboru; Wittbrodt, Joachim; Kondoh, Hisato

doi:10.1016/j.mod.2004.04.016

Cited by 127 publications

(85 citation statements)

References 62 publications

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“…Medaka is thought to be a model organism complimentary to zebrafish because it is evolutionarily distant to zebrafish and has a different organization of the genome, which is more similar to Fugu (reviewed in Kimura et al, 2004). In addition to completion of the medaka draft genome (Kasahara et al, 2007), a large number of medaka mutants has been isolated from natural population or from ethylnitrosourea (ENU) -induced mutagenesis screening (Ishikawa, 2000;Loosli et al, 2000;Furutani-Seiki et al, 2004; H.T., unpublished data; many of them can be browsed at National BioResource Project Medaka in Japan at http:// shigen.lab.nig.ac.jp/medaka). As expected, some of those mutants exhibit unique phenotypes that have not been covered by zebrafish mutants.…”

Section: Medaka (The Japanese Killifish;mentioning

confidence: 99%

Phenotypic analysis of a novel chordin mutant in medaka

Takashima

Shimada

Kobayashi

et al. 2007

Developmental Dynamics

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We have isolated and characterized a ventralized mutant in medaka (the Japanese killifish; Oryzias latipes), which turned out to have a mutation in the chordin gene. The mutant exhibits ventralization of the body axis, malformation of axial bones, over-bifurcation of yolk sac blood vessels, and laterality defects in internal organs. The mutant exhibits variability of phenotypes, depending on the culture temperature, from embryos with a slightly ventralized phenotype to those without any head and trunk structures. Taking

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Section: Medaka (The Japanese Killifish;mentioning

confidence: 99%

Phenotypic analysis of a novel chordin mutant in medaka

Takashima

Shimada

Kobayashi

et al. 2007

Developmental Dynamics

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“…Medaka is a small egg-laying freshwater fish that allows both genetic and embryological analyses and is one of the three vertebrate model organisms in which genome-wide phenotype-driven mutant screens were carried out 1 . Divergence of functional overlap of related genes between medaka and zebrafish allows identification of novel phenotypes that are unidentifiable in a single species 2 , thus medaka and zebrafish are complementary for genetic dissection of the vertebrate genome functions.…”

Section: Introductionmentioning

confidence: 99%

Dechorionation of Medaka Embryos and Cell Transplantation for the Generation of Chimeras

Porazinski¹,

Wang²,

Furutani-Seiki³

2010

JoVE

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Medaka is a small egg-laying freshwater fish that allows both genetic and embryological analyses and is one of the three vertebrate model organisms in which genome-wide phenotype-driven mutant screens were carried out 1 . Divergence of functional overlap of related genes between medaka and zebrafish allows identification of novel phenotypes that are unidentifiable in a single species 2 , thus medaka and zebrafish are complementary for genetic dissection of the vertebrate genome functions. Manipulation of medaka embryos, such as dechorionation, mounting embryos for imaging and cell transplantation, are key procedures to work on both medaka and zebrafish in a laboratory. Cell transplantation examines cell autonomy of medaka mutations. Chimeras are generated by transplanting labeled cells from donor embryos into unlabeled recipient embryos. Donor cells can be transplanted to specific areas of the recipient embryos based on the fate maps 3 so that clones from transplanted cells can be integrated in the tissue of interest during development. Due to the hard chorion and soft embryos, manipulation of medaka embryos is more involved than in zebrafish. In this video, we show detailed procedures to manipulate medaka embryos. Video LinkThe video component of this article can be found at https://www.jove.com/video/2055/ Protocol 1. Development of the embryos 1. When they are laid, eggs are clustered because of attachment filaments on the chorion. To let embryos develop normally, it is necessary to separate eggs. Tangle and cut attachment filaments by holding the attachment filaments with two forceps. 2. After unclustering, eggs are separated from feces and algae and transferred to fresh embryo medium at a maximum density of 40 eggs per 6cm Petri dish. 3. Medaka embryos develop slightly slower than zebrafish at 27°C. The timing of hatching is different between the two species; medaka embryos hatch from the chorion in 7 days and immediately start to swim and eat, whereas zebrafish embryos hatch in 2 days but start to swim and eat in 5-6 days. Development of medaka is staged according to Iwamatsu's staging 4 . 4. The development of medaka embryos can be conveniently adjusted to experimental plans by selecting the appropriate temperature.Development of medaka embryos can be stopped at 4°C in early development. After stage 24 when heart beating starts, development can be slowed using a minimum temperature of 18°C. 5. The timing of appearance of organs/tissues is slightly different in medaka compared with zebrafish, i.e. in medaka somitogenesis occurs after the onset of brain development whereas in zebrafish somitegenesis precedes brain development. Removing the chorionThe chorion of medaka consists of two protective layers with a hard inner layer and a soft outer surface. Thus, a two-step protease treatment employing pronase and hatching enzyme is necessary to remove this chorion.Once dechorionated, embryos should be kept in 1X BSS. Semi-sterile conditions will enhance the successful culture of dechorionated embryos, esp...

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“…Second, there are highly polymorphic inbred medaka strains available that can be used for mutagenesis screening and genetic mapping (Naruse et al 2004). These features significantly contribute to excellent forward genetics studies using medaka, such as the identification of the first sex-determining gene in non-mammalian vertebrates (Matsuda et al 2002(Matsuda et al , 2007, a large-scale mutagenesis screen to identify genes acting in diverse developmental processes (Furutani-Seiki et al 2004), and a fine quantitative trait locus analysis (Kimura et al 2007).…”

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Efficient Targeted Mutagenesis in Medaka Using Custom-Designed Transcription Activator-Like Effector Nucleases

et al. 2013

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Transcription activator-like effector nucleases (TALENs) have become powerful tools for targeted genome editing. Here we demonstrate efficient targeted mutagenesis in medaka (Oryzias latipes), which serves as an excellent vertebrate model for genetics and genomics. We designed and constructed a pair of TALENs targeting the medaka DJ-1 gene, a homolog of human DJ-1 (PARK7). These TALENs induced a number of insertions and deletions in the injected embryos with extremely high efficiency. This induction of mutations occurred in a dose-dependent manner. All screened G0 fish injected with the TALENs transmitted the TALEN-induced mutations to the next generation with high efficiency (44-100%). We also confirmed that these TALENs induced site-specific mutations because none of the mutations were found at potential off-target sites. In addition, the DJ-1 protein was lost in DJ-1 Δ7/Δ7 fish that carried a TALEN-induced frameshift mutation in both alleles. We also investigated the effect of the N-and C-terminal regions of the transcription activator-like (TAL) effector domain on the gene-disrupting activity of DJ1-TALENs and found that 287 amino acids at the N terminus and 63 amino acids at the C terminus of the TAL domain exhibited the highest disrupting activity in the injected embryos. Our results suggest that TALENs enable us to rapidly and efficiently establish knockout medaka strains. This is the first report of targeted mutagenesis in medaka using TALENs. The TALEN technology will expand the potential of medaka as a model system for genetics and genomics.

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A systematic genome-wide screen for mutations affecting organogenesis in Medaka, Oryzias latipes

Cited by 127 publications

References 62 publications

Phenotypic analysis of a novel chordin mutant in medaka

Phenotypic analysis of a novel chordin mutant in medaka

Dechorionation of Medaka Embryos and Cell Transplantation for the Generation of Chimeras

Efficient Targeted Mutagenesis in Medaka Using Custom-Designed Transcription Activator-Like Effector Nucleases

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