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在珠宝黄蜂中直接亲属 (DIPA) 的CRISPR, Nasonia vitripennis

Xinmi Zhang¹, Anabhra Singh¹, Kassandra Soriano Martinez¹

¹Department of Natural Sciences, Pitzer College and Scripps College, 925 N Mills Ave, Claremont, CA 91711, USA.

G3 (Bethesda, Md.)

|May 12, 2024

概括

直接父母 (DIPA-) CRISPR通过准眼睛色素基因,使珠宝黄蜂Nasonia vitripennis中的基因编辑成为可能. 优化注射参数对于一致的突变产生至关重要,尽管突变率仍然很低.

科学领域:

遗传学遗传学是一个
分子生物学分子生物学
昆虫学昆虫学是一门学科.

背景情况:

集群定期间隔的短平行体重复 (CRISPR) -Cas9技术提供了强大的基因编辑能力.
在某些昆虫物种中,有效的试剂传递仍然是CRISPR-Cas9在某些昆虫物种中的应用的挑战,包括珠宝黄蜂Nasonia vitripennis.
直接家长 (DIPA-) CRISPR提供了一种通过成人注射诱导基因病变的简化方法.

研究的目的:

为了评估DIPA-CRISPR在珠宝黄蜂Nasonia vitripennis中的疗效.
针对两个关键的眼睛色素基因,青菜和红,在欧莫色通路内.
优化Cas9-sgRNA度和注射量,以产生一致的突变.

主要方法:

通过准N. vitripennis. 中的青菜花和花基因来评估DIPA-CRISPR的功能.
系统地改变Cas9-sgRNA度和注射量,以确定最佳条件.
将N. vitripennis的突变率与其他昆虫物种中观察到的突变率进行比较.

主要成果:

成功生成了菜和花两种基因中的基因病变,证实了DIPA-CRISPR在N. vitripennis中的功能.
确定更大的注射量与各种试剂度相结合是产生一致突变的必要条件.
观察到与其他昆虫相比,N. vitripennis的总体突变率相对较低,可能是由于维特洛基因进口效率低.

结论:

DIPA-CRISPR是一种可行的基因编辑方法 Nasonia vitripennis,扩大其适用于更广泛的昆虫.
注射参数,特别是体积和试剂度,显著影响突变效率.
需要进行进一步的研究,以了解限制N. vitripennis突变率的因素,并将DIPA-CRISPR与替代传递方法进行比较.

关键词:

这种植物名为Nasonia vitripennis.迪帕迪帕是什么意思基因编辑基因编辑微注射微注射的方法生命的诞生 (vitellogenesis) 是因为

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