Hosoi Sayaka, Sakuma Tetsushi, Sakamoto Naoaki, Yamamoto Takashi
Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima, 739-8526, Japan.
Dev Growth Differ. 2014 Jan;56(1):92-7. doi: 10.1111/dgd.12099. Epub 2013 Nov 21.
Genome editing with engineered nucleases such as zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) has been reported in various animals. We previously described ZFN-mediated targeted mutagenesis and insertion of reporter genes in sea urchin embryos. In this study, we demonstrate that TALENs can induce mutagenesis at specific genomic loci of sea urchin embryos. Injection of TALEN mRNAs targeting the HpEts transcription factor into fertilized eggs resulted in the impairment of skeletogenesis. Sequence analyses of the mutations showed that deletions and/or insertions occurred at the HpEts target site in the TALEN mRNAs-injected embryos. The results suggest that targeted gene disruption using TALENs is feasible in sea urchin embryos.
利用锌指核酸酶(ZFNs)和转录激活样效应因子核酸酶(TALENs)等工程核酸酶进行基因组编辑已在多种动物中得到报道。我们之前描述了ZFN介导的海胆胚胎靶向诱变和报告基因插入。在本研究中,我们证明TALENs可在海胆胚胎的特定基因组位点诱导诱变。将靶向HpEts转录因子的TALEN mRNA注射到受精卵中导致骨骼生成受损。对突变的序列分析表明,在注射TALEN mRNA的胚胎中,HpEts靶位点发生了缺失和/或插入。结果表明,利用TALENs进行靶向基因破坏在海胆胚胎中是可行的。