An Editing-Site-Specific PCR Method for Detection and Quantification of CAO1-Edited Rice
- Resource Type
- Authors
- Shanshan Zhai; Yuhua Wu; Shengbo Zhao; Hongfei Gao; Xiaohong Yan; Yunjing Li; Nengwu Si; Jun Li; Fang Xiao; Zhang Hongwen; Gang Wu
- Source
- Foods, Vol 10, Iss 1209, p 1209 (2021)
Foods
Volume 10
Issue 6
- Subject
- 0106 biological sciences
Health (social science)
editing-site-specific PCR
Plant Science
Computational biology
TP1-1185
Biology
01 natural sciences
Health Professions (miscellaneous)
Microbiology
Article
03 medical and health sciences
Genome editing
Locked nucleic acid
Rice plant
CAO1-edited rice
030304 developmental biology
0303 health sciences
genome-edited plants
Chemical technology
fungi
food and beverages
quantification
Genetically modified organism
identification
Pcr method
Primer (molecular biology)
010606 plant biology & botany
Food Science
- Language
- English
- ISSN
- 2304-8158
Genome-edited plants created by genome editing technology have been approved for commercialization. Due to molecular characteristics that differ from classic genetically modified organisms (GMOs), establishing regulation-compliant analytical methods for identification and quantification of genome-edited plants has always been regarded as a challenging task. An editing-site-specific PCR method was developed based on the unique edited sequence in CAO1-edited rice plants. Test results of seven primer/probe sets indicated that this method can identify specific CAO1-edited rice from other CAO1-edited rice and wild types of rice with high specificity and sensitivity. The use of LNA (locked nucleic acid) in a probe can efficiently increase the specificity of the editing-site-specific PCR method at increased annealing temperature which can eliminate non-specific amplification of the non-target. The genome-edited ingredient content in blinded samples at the level of 0.1% to 5.0% was accurately quantified by this method on the ddPCR platform with RSD of <
15% and bias in the range of ±17%, meeting the performance requirements for GMO detection method. The developed editing-site-specific PCR method presents a promising detection and quantification technique for genome-edited plants with known edited sequence.