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However, negative correlation is also found between formation of some ecRNA and their mature linear RNA, possibly caused by competition between forward splicing and reverse splicing (or back splice).
Furthermore, the junction sites of the circRNA identified among various eukaryotic species are mostly flanked by canonical splice sites, the 5′ donor site GU and 3′ acceptor site AG, caused by a splicing event called the "back splice" (Jeck et al., 2013).
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Conserved RNAs formed by pre-mRNA back splicing, the function of which may be linked to that of their host genes.
If the mutant exon 2 is very efficiently spliced, there may be no substrate for back splicing.
Most circRNAs are derived from protein-coding pre-mRNAs, via a mechanism called "back-splicing".
We further discuss several important questions regarding the splicing of long noncoding RNAs and back-splicing of circular RNAs in cancers.
Existing methods of circRNA expression thus typically involve genes that have been engineered to contain sequence elements that promote back-splicing.
The more credible way to recognize circRNA for a specific gene is using PCR with out-facing primer pair (located near the back-splicing sites on the corresponding linear transcript), though false-positive cannot be excluded because of various reasons.
One could imagine that a deletion simply removes a specific sequence required by the splicing machinery to successfully back-splice.
As the authors state, a secondary structure would shorten the effective distance between the exon ends, but it may also possibly interfere with assembly of spliceosomal components or alter their bound conformation, which perhaps could positively affect the ability to back-splice.
This pathway predicts that back-splicing of exon 2 within the lariat will yield a doubly branched RNA product containing the two flanking introns.
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