Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Dev Cell. 2019 Mar 25;48(6):827-839.e9. doi: 10.1016/j.devcel.2019.01.020. Epub 2019 Feb 21.
The recent work of Besseling and Bringmann (2016) identified a molecular intervention for C. elegans in which premature segregation of maternal and paternal chromosomes in the fertilized oocyte can produce viable animals exhibiting a non-Mendelian inheritance pattern. Overexpression in embryos of a single protein regulating chromosome segregation (GPR-1) provides a germline derived clonally from a single parental gamete. We present a collection of strains and cytological assays to consistently generate and track non-Mendelian inheritance. These tools allow reproducible and high-frequency (>80%) production of non-Mendelian inheritance, the facile and simultaneous homozygosis for all nuclear chromosomes in a single generation, the precise exchange of nuclear and mitochondrial genomes between strains, and the assessments of non-canonical mitosis events. We show the utility of these strains by demonstrating a rapid assessment of cell lineage requirements (AB versus P1) for a set of genes (lin-2, lin-3, lin-12, and lin-31) with roles in C. elegans vulval development.
贝斯林和布林根(2016 年)的最新研究确定了一种针对秀丽隐杆线虫的分子干预方法,即在受精卵中母源和父源染色体过早分离可以产生具有非孟德尔遗传模式的可育动物。在胚胎中过量表达一种调节染色体分离的单一蛋白(GPR-1),可以从单个亲代配子中产生来源于生殖系的克隆。我们提供了一系列品系和细胞学检测方法,可稳定地产生和跟踪非孟德尔遗传。这些工具允许可重复且高频(>80%)地产生非孟德尔遗传,在单一代中方便且同时使所有核染色体纯合,在菌株之间精确交换核和线粒体基因组,并评估非典型有丝分裂事件。我们通过快速评估一组基因(lin-2、lin-3、lin-12 和 lin-31)在秀丽隐杆线虫的发育过程中对线粒体的需求(AB 与 P1),展示了这些品系的实用性。