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Transcriptome-wide off-target RNA editing induced by CRISPR-guided DNA base editors [P2A-EGFP control]

UID: 11040

Author(s): Grunewald, Julian, Lareau, Caleb A.* * MSK affiliated

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 or HepG2 cells were transfected with P2A-EGFP. Cells were sorted for top 5% GFP based on FITC signal. RNA-seq was performed to measure transcriptional changes associated with different constructs."
Subject of Study
Subject(s)
Access via GEO


Accession #: GSE129131

Access via BioProject


Accession #: PRJNA530210

Access via SRA


Accession #: SRP190024

Access Restrictions
Free to All
Access Instructions
The NCBI Gene Expression Omnibus. BioProject, and SRA databases provide open access to these files.
Associated Publications
Equipment Used
Illumina HiSeq 2500
Dataset Format(s)
TSV
Dataset Size
180.1 KB
Data Catalog Record Updated
2023-11-15