Extended Data Fig. 4: Screen for genes encoding PZM compartment components required for germline immortality and/or RNAi.
a, Introducing HA or GFP to the endogenous locus of newly identified Mrt genes using CRISPR/Cas9 did not abolish the protein product function. Lineages of animals of the indicated genotypes were grown in triplicate at 25 °C. ‘F’ indicates the generation at which one replicate lineage became sterile. As different lineages of the same mutant strain often become sterile on different generations, 1 to 3 'F' numbers are provided for each strain. For those generations where a lineage became sterile, brood sizes of control (WT, N2 Bristol) and animals harboring the corresponding epitope-tagged protein lineages were scored. Note, to minimize labor, brood sizes were only scored on the generation where a mutant lineage displayed sterility. n = 3 biological replicates, error bar, ± s.d. b, Fluorescence imaging of approximately a dozen germline cells in the pachytene region of WT or zsp-1(-) animals expressing GFP::MIP-2. Scale bar, 5 μm. n > 3 animals. c, Localization of F55A12.5. F55A12.5::HA was detected with an anti-HA antibody followed by Alex Fluor 568-conjugated secondary antibody. Scale bar, 20 μm for germline, 10 μm for embryos. n > 3 animals. d, CRISPR/Cas9-generated viable deletion mutants were subjected to pos-1 RNAi, and the percentage of hatched progeny was scored. Note, F01G4.4a.1 mutants exhibited an egg-lay defect phenotype; thus, the hatching rate was scored in utero. n = 3 biological replicates. Data are presented as mean ± s.d. Student’s t-test, two-tailed. P = 7.82 Ă— 10−5 (T26A5.2(ths364)), 3.72 Ă— 10−6 (mip-2(ths285)), 3 Ă— 10−5 (hpo-40(ths394)), 0.25 (herd-1(ths351)), 4.23 Ă— 10−7 (B0001.3(ths358)), 3.07 Ă— 10−10 (F52D2.12(ths438)), 1.48 Ă— 10−8 (pqn-53(ths427)), 0.0001 (C36B1.11(ths356)), 0.09 (F56C9.6(ths410)), 0.12 (B0261.7(ths367)), 1.79 Ă— 10−8(ubr-4(ths407)), 5.92 Ă— 10−5 (gln-3(ths387)), 0.0001 (H05C05.1(ths448)), 8.39 Ă— 10−5 (F59A3.12(ths520)), 5.12 Ă— 10−6 (ZK1307.1(ths597)), 1.52 Ă— 10−5 (B0001.8(ths352)), 0.051 (Y57A10A.31(ths272)), and 1.43 Ă— 10−5 (Y37H2A.1(ths603)). ns P ≥ 0.05, and **** P ≤ 0.0001. Source numerical data are available in source data.
