为了对基因组编辑产品进行精准定性和定量检测，以水稻SP1 基因的编辑植株为材料，在编辑位点上下
游设计通用引物，在编辑位点处设计基因编辑位点特异性TaqMan探针，建立了编辑位点特异性PCR方法。利用该
方法可准确鉴定特异基因组编辑产品，检测灵敏度达到5~10拷贝，可在实时荧光PCR（qPCR）和微滴数字PCR
（ddPCR）平台上对基因组编辑产品进行定量检测。由于数字PCR的微反应单元可消除野生型DNA对通用引物的
竞争性消耗，与qPCR的定量结果相比，ddPCR定量结果具有更高的定量准确性。

Genome editing technology has been successfully used in agricultural breeding, it is urgent to estab⁃
lish accurate identification and quantification technology based on molecular characteristics of genome-edited prod⁃
ucts. In this study, SP1 gene-edited rice was used as materials to develop an editing site-specific PCR method by
designing universal primer pair upstream and downstream of the editing sites, and gene editing site-specific Taq⁃
Man probes targeting at the editing sites. The results demonstrated that this method accurately identified specific ge⁃
nome-edited products with a detection sensitivity of 5-10 copies. The quantitative detection of genome-edited prod⁃
ucts was performed by both real-time PCR (qPCR) and droplet digital PCR (ddPCR). Since thousands of indepen⁃
dent droplets of ddPCR might eliminate the competitive consumption of universal primers by wild-type DNA, the
quantitative results of ddPCR showed higher quantitative accuracy than those of qPCR.

基因组编辑技术利用位点特异性核酸酶（sitespecific
nuclease, SSN）切割植物基因组DNA的特定
位点，产生双链断裂缺口（double stranded break,
DSB）[
1,2]，通过非同源末端连接（non-homologous end
joining, NHEJ）或同源重组（homologous recombina⁃
tion, HR）[
3]机制对双链断裂缺口进行修复，导致修复
位点发生单碱基或寡核苷酸片段的插入、缺失或替
换等突变，实现对基因组靶基因序列的精确编辑[4]。
基因组编辑常用的特异性核酸酶主要包括锌指核
酸酶（zinc finger nucleases, ZFNs）、类转录激活因子
效应物核酸酶（transcription activator-like effector
nucleases, TALENs）、以及规则簇状间隔的短回文重
复相关蛋白（clustered regularly interspaced short pal⁃
indromic repeat-associated protein，CRISPR/Cas）[
5]。
目前，基因组编辑技术已经成功用于多种植物的性
状改良，如增加千粒重的水稻[6]、低镉含量的籼稻[7]、
低糖含量的马铃薯[8]、糯性玉米[9]、高油酸大豆[10]、抗
溃疡病的柑橘[11]等；在北美洲，已经有两种基因组编
2021，43（1）：77-89
doi：
收稿日期：2020⁃11⁃03
基金项目：
be completed in 3 minutes on site, it was considered to be successfully meeting the requirements on site.