A simple method to dramatically increase C. elegans germline microinjection efficiency.
A simple method to dramatically increase C. elegans germline microinjection efficiency.
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一种显着提高秀丽隐杆线虫种系显微注射效率的简单方法。
DOI：
10.1101/2023.03.23.533855
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Porter,Rober
中科院分区：
文献类型：
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作者：
Gibney,TheresaV;Favichia,Michelle;Latifi,Laila;Medwig-Kinney,TaylorN;Matus,DavidQ;McIntyre,DanielC;Arrigo,AngeloB;Branham,KendallR;Bubrig,LouisT;Ghaddar,Abbas;Jiranek,JulianaA;Liu,KendraE;Marcucci,CharlesG;Porter,Rober
基因组操纵方法公司。雅典娜人需要将DNA或核糖核蛋白复合体显微注射到性腺合体的微观核心。这些微量注射在技术上要求很高，是所有基因组工程和转基因方法公司的关键瓶颈。优雅女装。虽然在遗传方法的简便性和效率方面有了稳步的改进。在基因组操作方面，在显微注射的物理过程中还没有类似的进展。在这里，我们报告了一种简单而廉价的方法，在注射过程中使用画笔处理蠕虫，与传统的蠕虫处理方法相比，平均微注射速度几乎增加了两倍。我们发现，通过显著提高注射速度和注射后存活率，画笔增加了注射吞吐量。除了显著和普遍地提高经验丰富的人员的注射效率外，画笔方法还显著提高了新手研究人员在微注射过程中执行关键步骤的能力。我们预计这一方法将使C。通过提高产生新菌株的速度，并将使基于显微注射的方法不那么具有挑战性，使没有广泛经验的人员和实验室更容易获得这种方法，从而使ELEAN更容易在社区内传播。
Genome manipulation methods inC. elegansrequire microinjecting DNA or ribonucleoprotein complexes into the microscopic core of the gonadal syncytium. These microinjections are technically demanding and represent a key bottleneck for all genome engineering and transgenic approaches inC. elegans. While there have been steady improvements in the ease and efficiency of genetic methods forC. elegansgenome manipulation, there have not been comparable advances in the physical process of microinjection. Here, we report a simple and inexpensive method for handling worms using a paintbrush during the injection process that nearly tripled average microinjection rates compared to traditional worm handling methods. We found that the paintbrush increased injection throughput by substantially increasing both injection speeds and post-injection survival rates. In addition to dramatically and universally increasing injection efficiency for experienced personnel, the paintbrush method also significantly improved the abilities of novice investigators to perform key steps in the microinjection process. We expect that this method will benefit theC. eleganscommunity by increasing the speed at which new strains can be generated and will also make microinjection-based approaches less challenging and more accessible to personnel and labs without extensive experience.