嗜卵磷脂海胆赤藓的胚胎显微注射

A. Edgar, M. Byrne, G. Wray
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引用次数: 3

摘要

微注射是一种常见的胚胎学技术,用于许多类型的实验,包括谱系追踪、操纵基因表达或基因组编辑。可注射试剂包括检测感兴趣基因的信使核糖核酸过表达、错误表达或显性阴性实验、防止感兴趣信使核糖核酸或剪接形式翻译的吗啉反义寡核苷酸和CRISPR-Cas9试剂。因此,该技术可广泛用于基础胚胎学研究、构建基因调控网络以及直接测试发育进化过程中顺式调控和编码序列变化的假设。然而,在典型的浮游海洋无脊椎动物中进行显微注射的方法可能在卵磷脂营养物种的高度修饰的卵子和胚胎中效果不佳。该方案针对嗜卵磷脂海胆Heliocidaris赤眼蜂进行了优化。
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Embryo microinjection of the lecithotrophic sea urchin Heliocidaris erythrogramma
Microinjection is a common embryological technique used for many types of experiments, including lineage tracing, manipulating gene expression, or genome editing. Injectable reagents include mRNA overexpression, mis-expression, or dominant-negative experiments to examine a gene of interest, a morpholino antisense oligo to prevent translation of an mRNA or spliceoform of interest and CRISPR-Cas9 reagents. Thus, the technique is broadly useful for basic embryological studies, constructing gene regulatory networks, and directly testing hypotheses about cis-regulatory and coding sequence changes underlying the evolution of development. However, the methods for microinjection in typical planktotrophic marine invertebrates may not work well in the highly modified eggs and embryos of lecithotrophic species. This protocol is optimized for the lecithotrophic sea urchin Heliocidaris erythrogramma.
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