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Alignment to a unique genome position was enforced, effectively eliminating sequences derived from repeated elements.
When more than 20% of the contig length resulted from repeated elements, contigs were discarded.
Since the crasiRNAs are derived largely from repeated elements, it would be interesting to explore enrichment of discontiguous palindromic motifs in specific regions of the genome such as those enriched in repetitive elements and centromeric regions.
For example, a 1 kb bin that corresponds to a repeated sequence could be over represented because of the faster renaturation kinetics of repeated sequences, or it could be underrepresented if reads from repeated elements were removed by the algorithm used to map the reads (in our case Repeat Masker).
Transcription from repeated elements within these regions causes the formation of double stranded RNAs (dsRNAs), which are processed by Dcr1 to produce small interfering RNAs of 21 22 nucleotides (siRNAs) that are loaded onto the RNA-induced transcriptional silencing (RITS) complex.
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This analysis indicates the crasiRNAs are enriched for repeated elements (LINEs, SINEs, transposons), although it was not possible to determine from this mapping scheme whether the repeat elements themselves were associated with centromere domains.
Moreover, small RNAs derived from highly repeated elements were annotated as repeat-associated small RNAs.
Another fraction was derived from highly repeated elements named repeat-associated small RNAs.
To avoid noise from these repeated elements, a repeat masked reference genome was used.
The amount of generated sequence data is large enough to include multiple reads from highly repeated elements, thus allowing evaluation of their abundance and sequence composition.
We did not observe a significant enrichment of antisense reads in these genes (n = 198) in the nucleus against both the total and the cytosolic fractions at 20 h p.i., while ncRNA from telomere-associated repeated elements were enriched in the nuclear fraction (Additional file 7: Figure S4).
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