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ARS Home » Midwest Area » Columbia, Missouri » Plant Genetics Research » Research » Research Project #442556

Research Project: Development of Soybean Lines Without Both Trypsin and Chymotrypsin Inhibitors

Location: Plant Genetics Research

Project Number: 5070-21000-044-014-R
Project Type: Reimbursable Cooperative Agreement

Start Date: Oct 1, 2022
End Date: Mar 31, 2024

Objective:
Development of adapted soybean germplasm which bears drastically reduced (ideally eliminated) anti-nutritional Kunitz-trypsin inhibitors and Bowman-Birk inhibitors.

Approach:
We will employ gene editing (CRISPR CAS9) to delete an abundant trypsin inhibitor gene expressed in seeds. The transgenic events generated by the CRISPR CAS9 system will be planted in a greenhouse and seeds harvested from these plants will be utilized for measuring the trypsin and chymotrypsin inhibitor activities following well-established protocols. We have raised antibodies against soybean these inhibitors which will be used to quantify the amount of these proteins and activities in transgenic events by western blot analysis and/or ELISA. Non-transformed soybean plants will be used as controls. Gene edited transgenic soybeans will be crossed with each other to create a soybean line that lacks both trypsin and chymotrypsin inhibitors.