In Vitro CRISPR-Cpf1 Assay for Differentiation of Fine and Bulk Cocoa (Theobroma cacao L.)

Link:
Autor/in:
Erscheinungsjahr:
2022
Medientyp:
Text
Schlagworte:
  • Theobroma cacao L.
  • in vitro CRISPR-Cpf1
  • Cas12a
  • food fraud
  • Arriba
  • CCN-51
Beschreibung:
  • Cocoa cultivation is dominated by the clone “Colleción Castro Naranjal 51” (CCN-51). In contrast, CCN-51 is the expensive and aromatic fine cocoa “Arriba Nacional” from Ecuador. The differences in the overall quality of the beans and in the prices show that it is necessary to develop a rapid and accurate method to distinguish these varieties and prevent food fraud. To this end, we used a CRISPR-Cpf1 assay suitable for AT-rich targets such as the chloroplast genome (cpGenome). SNPs in cocoa plastid genomes were selected to replace the canonical PAM sequence of Cpf1 (5′-TTTV-3′). We developed two assay systems to digest both Arriba and CCN-51. The results were tested qualitatively by agarose gel electrophoresis and quantitatively by capillary gel electrophoresis. Using the assay described here, we were able to reliably detect admixtures of 5% CCN-51 (P < 0.01) and 10% Arriba (P < 0.05). The application to processed cocoa products was also successful.
Lizenz:
  • info:eu-repo/semantics/closedAccess
Quellsystem:
Forschungsinformationssystem der UHH

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oai:www.edit.fis.uni-hamburg.de:publications/a2e3afac-46ca-4d94-b2ac-2cae1d8a1129