通过注射 I-SceI 核酸内切酶或 Tol2 转座子系统进行蝾螈转基因。

Axolotl Transgenesis via Injection of I-SceI Meganuclease or Tol2 Transposon System.

机构信息

Center for Regenerative Therapies Dresden (CRTD), Technische Universität Dresden, Dresden, Germany.

Medical Faculty: Department of Internal Medicine 3, Center for Healthy Aging, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.

出版信息

Methods Mol Biol. 2023;2562:321-333. doi: 10.1007/978-1-0716-2659-7_21.

DOI:10.1007/978-1-0716-2659-7_21

PMID:36272085

Abstract

The axolotl (Ambystoma mexicanum ) has been widely used as an animal model for studying development and regeneration. In recent decades, the use of genetic engineering to alter gene expression has advanced our knowledge on the fundamental molecular and cellular mechanisms, pointing us to potential therapeutic targets. We present a detailed, step-by-step protocol for axolotl transgenesis using either I-SceI meganuclease or the mini Tol2 transposon system, by injection of purified DNA into one-cell stage eggs. We add useful tips on the site of injection and the viability of the eggs.

摘要

美西螈（Ambystoma mexicanum）被广泛用作研究发育和再生的动物模型。在过去几十年中，利用基因工程改变基因表达水平的方法极大地促进了我们对基本分子和细胞机制的认识，为潜在的治疗靶点指明了方向。本研究提供了详细的一步一步的protocol，通过将纯化的 DNA 注射到单细胞期的卵中，使用 I-SceI 核酸内切酶或 mini Tol2 转座子系统来实现美西螈的转基因。我们还添加了一些关于注射部位和卵活力的有用提示。