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Targeted mutagenesis using CRISPR-Cas9 in sea urchin, Hemicentrotus pulcherrimus

Liu Daming 広島大学

2020.03.03

概要

Sea urchins are invertebrate deuterostomes, which is a sister group of the chordates.
Investigation of the sea urchin genome is important to understand the origin of vertebrate
gene functions. Sea urchins are a model organism for research on developmental biology.
The fertilized eggs and embryos of sea urchin can be manipulated directly because of its
simple organization. It is therefore very suitable for studies of developmental process
such as morphogenetic movements, cell interactions, changes in gene expression
associated with the establishment of tissue territories (Becker et al., 2003). Furthermore,
sea urchin has been recently used for the analysis of gene regulatory networks during
early development (Davidson et al., 2002; Oliveri & Davidson, 2004; Oliveri et al.,
2008).
Although deuterostomes generally have bilateral bodies, the bilateral body plan of
sea urchin embryos and larvae is converted to a pentameral body plan after
metamorphosis. The metamorphosis of sea urchin is a complex developmental
progression, much of which takes place in the adult rudiment produced from the left side
of the coelomic pouch that originates from small micromere and macromere descendants
(Cameron et al., 1991; Cameron et al., 1987).
The first sea urchin genome sequenced was that of Strongylocentrotus purpuratus
(Sea Urchin Genome Sequencing Consortium, 2006). Hemicentrotus pulcherrimus,
which is closely related to S. purpuratus, is widely distributed in Japan and has also been
used for research on developmental biology. The H. pulcherrimus genome was
deciphered in 2018 (Kinjo et al., 2018). The genome size of H. pulcherrimus was
estimated to be 800 Mbp, with approximately 25,000 genes.
Analysis of gene function in the sea urchin embryo has relied on gene knockdown
by morpholino antisense oligonucleotide (MASO), which blocks translation or RNA
splicing of gene transcripts of interest (Angerer & Angerer, 2004). However, the effect
of MASO‐mediated knockdown is not long lasting. Therefore, it is impossible to perturb
gene expression during late developmental stages by MASO. ...

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Targeted mutagenesis using CRISPR-Cas9 in sea urchin, Hemicentrotus pulcherrimus

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