Quantitative Cell Biology, Rhineland-Palatinate Technical University, Kaiserslautern, Germany.
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
Methods Enzymol. 2024;706:97-123. doi: 10.1016/bs.mie.2024.07.022. Epub 2024 Aug 15.
Budding yeast is a laboratory model of a simple eukaryotic cell. Its compact genome is very easy to edit. This allowed to create systematic collections (libraries) of yeast strains where every gene is either perturbed or tagged. Here we review how such collections were used to study mitochondrial biology by doing genetic screens. First, we introduce the principles of yeast genome editing and the basics of its life cycle that are useful for genetic experiments. Then we overview what yeast strain collections were created over the past years. We also describe the creation and the usage of the new generation of SWAP-Tag (SWAT) collections that allow to create custom libraries. We outline the principles of changing the genetic background of whole collections in parallel, and the basics of synthetic genetic array (SGA) approach. Then we review the discoveries that were made using different types of genetic screens focusing on general mitochondrial functions, proteome, and protein targeting pathways. The development of new collections and screening techniques will continue to bring valuable insight into the function of mitochondria and other organelles.
芽殖酵母是一种简单真核细胞的实验室模型。其紧凑的基因组非常易于编辑。这使得可以创建系统的酵母菌株集合（文库），其中每个基因都受到干扰或标记。在这里，我们将通过遗传筛选来回顾这些集合如何用于研究线粒体生物学。首先，我们介绍了酵母基因组编辑的原理和其生命周期的基础知识，这些对于遗传实验非常有用。然后，我们概述了过去几年中创建的酵母菌株集合。我们还描述了创建和使用新的 SWAP-Tag（SWAT）集合的方法，该集合允许创建自定义文库。我们概述了并行更改整个集合遗传背景的原则，以及合成遗传阵列（SGA）方法的基础知识。然后，我们回顾了使用不同类型的遗传筛选方法发现的结果，这些方法主要集中在一般线粒体功能、蛋白质组和蛋白质靶向途径上。新集合和筛选技术的发展将继续为线粒体和其他细胞器的功能提供有价值的见解。
