Department of Biological Chemistry, School of Medicine, University of California Irvine, Irvine, CA 92697, USA.
Chao Family Comprehensive Cancer Center, University of California Irvine, Irvine, CA 92697, USA.
STAR Protoc. 2022 Mar 9;3(1):101148. doi: 10.1016/j.xpro.2022.101148. eCollection 2022 Mar 18.
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).
APOBEC3A、CRISPR 可编程 RNA 碱基编辑器或其他酶可以在特定位置或热点编辑 RNA 转录本。精确量化这些 RNA 编辑事件对于确定这些酶在细胞中的活性和效率至关重要。我们开发了一种使用数字 PCR 快速定量 RNA 编辑活性的方法，该技术通过微分区批量 PCR 反应来灵敏且定量地检测稀有突变。该测定法可快速定量细胞系或患者样本中的 RNA 编辑事件。有关该方案的使用和执行的完整详细信息，请参阅 Jalili 等人（2020 年）和 Oh 等人（2021 年）。
