Jhuang Han-Ying, Aggeli Dimitra, Lang Gregory I
Department of Biological Sciences, Lehigh University, Bethlehem, PA, USA.
Yeast. 2025 Jun;42(5-7):126-131. doi: 10.1002/yea.4001. Epub 2025 Apr 7.
Introducing plasmids into yeast is a critical step for many phenotypic assays and genetic engineering applications. However, it is often challenging for applications that involve large pools of variants because the population structure can be easily altered by traditional methods such as chemical transformation. In this study, we introduce drug-marked plasmids into a heterogeneous yeast population using both transformation and cytoduction (mating without nuclear fusion). Using a highly diverse barcoded yeast collection, we quantify the efficiency of both methods. We demonstrate that for cytoduction, but not transformation, nearly all the genotypes in the initial pool were detected in the final pool, with a high correlation to their initial frequencies. Finally, we map QTL that impact both cytoduction and transformation. Overall, we demonstrate the efficiency of cytoduction as a means of introducing plasmids into yeast. This is significant because it provides a means of manipulating diverse yeast populations, such as pools constructed for bulk segregant analysis, deep mutational scanning, large-scale gene editing, or populations from long-term evolution experiments.
将质粒导入酵母是许多表型分析和基因工程应用的关键步骤。然而，对于涉及大量变异体的应用来说，这通常具有挑战性，因为群体结构很容易被化学转化等传统方法改变。在本研究中，我们通过转化和细胞融合（无核融合的交配）将带有药物标记的质粒导入异质酵母群体。使用高度多样化的条形码酵母文库，我们对这两种方法的效率进行了量化。我们证明，对于细胞融合而非转化，初始库中的几乎所有基因型在最终库中都能被检测到，且与它们的初始频率高度相关。最后，我们定位了影响细胞融合和转化的数量性状基因座。总体而言，我们证明了细胞融合作为将质粒导入酵母的一种方法的效率。这具有重要意义，因为它提供了一种操纵多样化酵母群体的方法，例如为大量分离分析、深度突变扫描、大规模基因编辑构建的文库，或来自长期进化实验的群体。
