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While two deletion alleles exist for tdp-1, tdp-1 ok803) tdp-1 ok803k781), only the tdp-1(ok803) andele tdp-1 ok781 tdp-1 ok781null (Supplementary Fig S1B and C); therefonly our sthey tdp-1 ok803 tdp-1 ok803le.
As shown in Supplementary Fig S15, the RIP experiments confirmed TDP-1's association with a subset of introns that were bound by TDP-1 in the ChIP-seq.
20 μl TDP-1 antibody was used.
Analysis of TDP-1 binding by deep sequencing of anti-TDP-1 chromatin immunoprecipitation (ChIP) indicated that TDP-1 associates with highly structured regions co-transcriptionally.
These same transcripts are associated with TDP-1 and contain increased amounts of dsRNA structure/abundance in tdp-1 mutants.
To determine whether TDP-1 acts directly on transcripts with potential double-stranded structure, we asked whether TDP-1 protein associates with genes containing excessive dsRNA in tdp-1 ok803 tdp-1 ok803
Analysis of TDP-1 binding in genes with increased dsRNA structure/stability in tdp-1 mutants (as assayed by J2-IP enrichment) showed about 40% of these genes contained clear TDP-1 binding sites (P = 1 × 10−21, hgd).
While splicing abnormalities may contribute to tdp-1 loss of function defects, splicing differences do not readily explain changes in transcript abundance in tdp-1 ok803 tdp-1 ok803
Despite the lack of severe phenotypes in the tdp-1 mutant, TDP-1 has molecular properties similar to its mammalian homolog.
These results imply that while tdp-1 loss of function may be less consequential in the worm, TDP-1's basic molecular roles are likely conserved.
Because deletion of tdp-1 perturbed the abundance of these transcripts, we hypothesized that TDP-1 has a fundamental function in the formation, or metabolism, of dsRNA.
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