利用 CRISPR/Cas9 和 TALEN 技术提高海鞘基因组编辑效率。

Improved Genome Editing in the Ascidian Ciona with CRISPR/Cas9 and TALEN.

机构信息

Shimoda Marine Research Center, University of Tsukuba, Shimoda, Shizuoka, Japan.

Laboratory for Single-Cell Neurobiology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan.

出版信息

Methods Mol Biol. 2023;2637:375-388. doi: 10.1007/978-1-0716-3016-7_28.

DOI:10.1007/978-1-0716-3016-7_28

PMID:36773161

Abstract

The ascidian Ciona intestinalis type A (or Ciona robusta) is an important organism for elucidating the mechanisms that make the chordate body plan. CRISPR/Cas9 and TAL effector nuclease (TALEN) are widely used to quickly address genetic functions in Ciona. Our previously reported method of CRISPR/Cas9-mediated mutagenesis in this animal has inferior mutation rates compared to those of TALENs. We here describe an updated way to effectively mutate genes with CRISPR/Cas9 in Ciona. Although the construction of TALENs is much more laborious than that of CRISPR/Cas9, this technique is useful for tissue-specific knockouts that are not easy even by the optimized CRISPR/Cas9 method.

摘要

海鞘 Ciona intestinalis 型 A（或 Ciona robusta）是阐明使脊索动物体模式形成的机制的重要生物体。CRISPR/Cas9 和 TAL 效应物核酸酶（TALEN）被广泛用于快速解决 Ciona 中的遗传功能。我们之前报道的这种动物中的 CRISPR/Cas9 介导的诱变方法与 TALEN 的突变率相比要低。我们在这里描述了一种用 CRISPR/Cas9 有效突变 Ciona 中基因的更新方法。虽然 TALEN 的构建比 CRISPR/Cas9 复杂得多，但该技术对于组织特异性敲除很有用，即使通过优化的 CRISPR/Cas9 方法也不容易实现。