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A digital PCR-based protocol to detect and quantify RNA editing events at hotspots.

ABSTRACT: APOBEC3A, CRISPR programmable RNA base editors, or other enzymes can edit RNA transcripts at specific locations or hotspots. Precise quantification of these RNA-editing events is crucial to determine the activity and efficiency of these enzymes in cells. We have developed a quick method to quantify RNA-editing activity using digital PCR, a sensitive and quantitative technique to detect rare mutations by micro-partitioning bulk PCR reactions. This assay allows rapid absolute quantification of RNA editing events in cell lines or patient samples. For complete details on the use and execution of this protocol, please refer to Jalili et al. (2020) and Oh et al. (2021).

SUBMITTER: Oh S

PROVIDER: S-EPMC8915000 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature

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A digital PCR-based protocol to detect and quantify RNA editing events at hotspots.

Oh Sunwoo S Buisson Rémi R

STAR protocols 20220309 1

APOBEC3A, CRISPR programmable RNA base editors, or other enzymes can edit RNA transcripts at specific locations or hotspots. Precise quantification of these RNA-editing events is crucial to determine the activity and efficiency of these enzymes in cells. We have developed a quick method to quantify RNA-editing activity using digital PCR, a sensitive and quantitative technique to detect rare mutations by micro-partitioning bulk PCR reactions. This assay allows rapid absolute quantification of RNA ...[more]

PMID: 35284835

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