Biallelic genome modification in F(0) Xenopus tropicalis embryos using the CRISPR/Cas system.

Biallelic genome modification in F(0) Xenopus tropicalis embryos using the CRISPR/Cas system.
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DOI:
10.1002/dvg.22719
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发表时间:
2013-12
期刊:
影响因子:
1.5
通讯作者:
Cho, Ken W. Y.
Cho, Ken W. Y.
中科院分区:
生物学4区
文献类型:
--
作者:
Blitz, Ira L.;Biesinger, Jacob;Xie, Xiaohui;Cho, Ken W. Y.

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基因失活是表型和基因组数据关联的重要工具,当基因功能受损时,研究人员可以根据表型推断正常的基因功能。需要新的和更好的方法来克服现有方法的不足,以显著加快科学进步。我们已经将CRISPR/Cas系统用于热带非洲爪哇,并报告了编码酪氨酸酶基因的有效突变,酪氨酸酶是导致眼皮肤白化的原因。在F0代检测到该基因的双等位突变,提示其靶向效率与TALEN相似。我们还发现,非靶标突变似乎可以忽略不计,因此CRISPR/Cas可能是一个有用的系统,在这个重要的模式生物中创造基因组修改。
Gene inactivation is an important tool for correlation of phenotypic and genomic data, allowing researchers to infer normal gene function based on the phenotype when the gene function is impaired. New and better approaches are needed to overcome the shortfalls of existing methods for any significant acceleration of scientific progress. We have adapted the CRISPR/Cas system for use in Xenopus tropicalis and report on the efficient creation of mutations in the gene encoding the enzyme tyrosinase, which is responsible for oculocutaneous albinism. Biallelic mutation of this gene was detected in the F0 generation, suggesting targeting efficiencies similar to that of TALENs. We also find that off-target mutagenesis appears to be negligible and therefore CRISPR/Cas may be a useful system for creating genome modifications in this important model organism.
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