Development of an Event-Specific Droplet Digital PCR Assay for Quantification and Evaluation of the Transgene DNAs in Trace Samples of GM -Knockout Goat

Overview

Journal Foods

Specialty Biotechnology

Date 2022 Mar 25

PMID 35327291

Authors

Wenting Xu

Ping Shen

Rong Li

Biao Liu

Litao Yang

Affiliations

Soon will be listed here.

Abstract

The prion protein () gene encoding prion protein is considered a prerequisite for the occurrence of scrapie disease, and knockout of the gene in transgenic goat is one effective approach to avoid scrapie. This study aims to establish an event-specific droplet digital polymerase chain reaction (ddPCR) assay to detect and quantify the content of genetically modified (GM) -knockout goat event KoP1. The developed ddPCR assay presents high specificity, sensitivity, accuracy, precision and wide dynamic range. The limits of detection and quantification were as low as 1.44 and 7.2 haploid genome equivalent (HGE) per reaction, respectively. Furthermore, this assay was successfully applied in quantifying the goat KoP1 GM content in milk, feces and living environmental soil samples. We believe that the developed ddPCR assay has the potential to be used in the evaluation of horizontal gene transfer and the practical risk assessment of GM goat event KoP1 and its derivatives.

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