Homo sapiens
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA316460 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR3305250.fastq.gz | Fastqsanger.gz | |||
| SRR3305260.fastq.gz | Fastqsanger.gz | |||
| SRR3305270.fastq.gz | Fastqsanger.gz | |||
| SRR3305280.fastq.gz | Fastqsanger.gz | |||
| SRR3305290.fastq.gz | Fastqsanger.gz |
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Project description:Base Editing has been touted the most intelligent and precise application of the CRISPR platform so far, merging the simplicity of RNA-guided nucleases with deaminases that allow for the programmable generation of single base substitutions - without introduction of double-strand breaks. Even though the two-component system has been expected to cause off-target substitutions, studies involving cytosine base editors (CBEs) showed that in most cases, relatively few single base off-targets could be detected on DNA. We introduce the concept of multi-dimensional off-targeting, presenting an extensive amount of RNA cytidines being edited by DNA base editors. Epitranscriptomic off-target effects affected different cell lines and were independent of the guide RNAs used, suggesting Cas9-independent activity of the cytidine deaminase rAPOBEC1 on single-stranded RNA. With the help of protein engineering, we developed CBE variants with massively reduced inadvertent mutation of RNA that preserve and enhance DNA base editing capabilities.
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