Assessment and Maintenance of Unigametic Germline Inheritance for C. elegans

• Published in: Developmental cell 2019-03, Vol.48 (6), p.827-839.e9
• Main Authors: Artiles, Karen L., Fire, Andrew Z., Frøkjær-Jensen, Christian
• Format: Article
• Language: English
• Subjects: Animals; Biomarkers - metabolism; C. elegans; Caenorhabditis elegans - embryology; Caenorhabditis elegans - genetics; Caenorhabditis elegans Proteins - genetics; Caenorhabditis elegans Proteins - metabolism; Chromosomes - genetics; Crosses, Genetic; Female; Fluorescence; genetic engineering; Germ Cells - metabolism; inheritance; Inheritance Patterns - genetics; Male; Microtubules - metabolism; mitosis; mosaic; Mosaicism; Mutation - genetics; non-Mendelian; Pharynx - metabolism; Phenotype; synthetic biology; Vulva - embryology; Vulva - metabolism; Zygote - metabolism
• ISSN: 1534-5807; 1878-1551
• DOI: 10.1016/j.devcel.2019.01.020

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. [Display omitted] •Non-Mendelian inheritance can be stably maintained using optimized GPR-1 transgenes•Fluorescently marked toolkit strains enable precise tracking of inheritance mode•Marking allows derivation of sperm-derived clonal populations from any fertile male•Toolkit enables precise tests for lineage-of-action and non-nuclear inheritance Non-Mendelian inheritance in C. elegans results in chimeric animals with cells containing only maternal or paternal nuclear DNA. Artiles et al. develop a toolkit of strains with a high frequency of non-Mendelian segregation and monitor inheritance by fluorescence, thereby enabling experiments requiring detection of non-canonical inheritance events across large populations.