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Image Search Results
Journal: bioRxiv
Article Title: An egg sabotaging mechanism drives non-Mendelian transmission in mice
doi: 10.1101/2024.02.22.581453
Figure Lengend Snippet: ( A ) R2d2 heterozygous (+/–) oocytes expressing H2B-mCherry, dCas9-EGFP, and gRNA targeting the R2d2 DNA sequence were imaged live at anaphase I (white line, oocyte cortex). The histogram shows the distribution of the fraction of lagging chromosomes that have dCas9 signals (n = 35 cells). For the non-recombinant chromosome example (top), we imaged the dCas9-EGFP signals only at the first timepoint and at the anaphase onset to minimize photobleaching and phototoxicity. In the recombinant chromosome example (bottom), three dCas9 signals were observed at the 2 hr 20 min timepoint due to the splitting of the R2d2 locus on one of the homologs (also see B). ( B ) R2d2 heterozygous (+/–) oocytes expressing H2B-mCherry, dCas9-EGFP, and gRNA targeting the R2d2 DNA sequence were imaged live at the metaphase I to anaphase I transition. The fraction of oocytes with the dCas9 signal splitting or stretching was quantified in metaphase I and anaphase I (n = 26 cells); Chi-Square Test of Independence was used to calculate the P values in the graph; white line, oocyte cortex; orange asterisks, stretching R2d2 locus. We used the Denoise.ai software (Nikon) to reduce noise and follow the R2d2 locus better in anaphase I.
Article Snippet: Due to the weak dCas9-GFP signals during live-imaging experiments , we used the
Techniques: Expressing, Sequencing, Recombinant, Software