Transcriptome-wide off-target RNA editing induced by CRISPR-guided DNA base editors [Modifications - screen]
UID: 11043
- Description
- Summary from GEO:
"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."
Overall design from GEO:
"HEK293T cells were transfected with pCAG-BE3-P2A-EGFP or variants thereof or control pCAG-nCas9(D10A)-UGI-NLS-P2A-EGFP or control pCAG-P2A-EGFP constructs with various gRNAs as described below. Cells were sorted for top 5% GFP or all GFP + cells based on FITC signal. RNA-seq was performed to measure transcriptional changes associated with different constructs and guides."
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Access via GEO
Accession #: GSE121666Access via BioProject
Accession #: PRJNA498065Access via SRA
Accession #: SRP166457 - Access Restrictions
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Free to All
- Access Instructions
- The NCBI Gene Expression Omnibus. BioProject, and SRA databases provide open access to these files.
- Associated Publications
- Equipment Used
- Dataset Format(s)
- TSV
- Dataset Size
- 369.4 KB
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