title: Molecular Characterization of Transgenic Events Using Next Generation Sequencing Approach.
abstract: Demand for the commercial use of genetically modified (GM) crops has been increasing in light of the projected growth of world population to nine billion by 2050. A prerequisite of paramount importance for regulatory submissions is the rigorous safety assessment of GM crops. One of the components of safety assessment is molecular characterization at DNA level which helps to determine the copy number, integrity and stability of a transgene; characterize the integration site within a host genome; and confirm the absence of vector DNA. Historically, molecular characterization has been carried out using Southern blot analysis coupled with Sanger sequencing. While this is a robust approach to characterize the transgenic crops, it is both time- and resource-consuming. The emergence of next-generation sequencing (NGS) technologies has provided highly sensitive and cost- and labor-effective alternative for molecular characterization compared to traditional Southern blot analysis. Herein, we have demonstrated the successful application of both whole genome sequencing and target capture sequencing approaches for the characterization of single and stacked transgenic events and compared the results and inferences with traditional method with respect to key criteria required for regulatory submissions.
authors.affiliation: Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, Indiana, 46268, United States of America.
authors.forename: Satish K
authors.initials: SK
authors.lastname: Guttikonda
authors.affiliation: Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, Indiana, 46268, United States of America.
authors.forename: Pradeep
authors.initials: P
authors.lastname: Marri
authors.affiliation: Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, Indiana, 46268, United States of America.
authors.forename: Jafar
authors.initials: J
authors.lastname: Mammadov
authors.affiliation: Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, Indiana, 46268, United States of America.
authors.forename: Liang
authors.initials: L
authors.lastname: Ye
authors.affiliation: Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, Indiana, 46268, United States of America.
authors.forename: Khaing
authors.initials: K
authors.lastname: Soe
authors.affiliation: Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, Indiana, 46268, United States of America.
authors.forename: Kimberly
authors.initials: K
authors.lastname: Richey
authors.affiliation: Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, Indiana, 46268, United States of America.
authors.forename: James
authors.initials: J
authors.lastname: Cruse
authors.affiliation: Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, Indiana, 46268, United States of America.
authors.forename: Meibao
authors.initials: M
authors.lastname: Zhuang
authors.affiliation: Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, Indiana, 46268, United States of America.
authors.forename: Zhifang
authors.initials: Z
authors.lastname: Gao
authors.affiliation: Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, Indiana, 46268, United States of America.
authors.forename: Clive
authors.initials: C
authors.lastname: Evans
authors.affiliation: Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, Indiana, 46268, United States of America.
authors.forename: Steve
authors.initials: S
authors.lastname: Rounsley
authors.affiliation: Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, Indiana, 46268, United States of America.
authors.forename: Siva P
authors.initials: SP
authors.lastname: Kumpatla
mesh_terms: D015139:Blotting, Southern; D018628:Gene Dosage; D023281:Genomics / Q000379:methods; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D000069600:Plant Breeding; D030821:Plants, Genetically Modified / Q000235:genetics*; D013025:Glycine max / Q000235:genetics; D019076:Transgenes