Open AccessMolecular Characterization of Transgenic Events Using Next Generation Sequencing Approach
Author(s) -
Satish K. Guttikonda,
Pradeep Reddy Marri,
Jafar Mammadov,
Y. T. Liang,
Khaing Soe,
Kimberly Richey,
James Cruse,
Meibao Zhuang,
Zili Gao,
Clive Evans,
Steve Rounsley,
Siva P. Kumpatla
Publication year - 2016
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0149515
Subject(s) - sanger sequencing , dna sequencing , computational biology , biology , genome , transgene , population , genetically modified organism , genetically modified crops , genetics , microbiology and biotechnology , dna , gene , medicine , environmental health
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.