CRISPR-Cas9 Mediates Targeted Gene Replacement and Knockin in ArabidopsisAugust 29, 2018
Homology-directed gene repair (HDR), in which the gene of interest is knocked in to the genome or replaced to an undesired DNA sequence, is a powerful tool in CRISPR-Cas9-mediated genome editing. However, this technique has limited successes in higher plants, although it has been applied to other organisms such as yeast, mice, and fruit fly.
Scientists from Shanghai Center for Plant Stress Biology and Center for Excellence in Molecular Plant Sciences developed a "sequential transformation" gene-targeting strategy to target endogenous DNA glycosylase genes ROS1 and DME in Arabidopsis. They first tested the applicability of the method by knocking in ROS1-GFP gene and found that the strategy can result to a stably inherited genomic locus as large as 1.6 kb.
To further test their method, they knocked in GFP at the 5′ end and the 3′ end of DME and found that the gene was stably transferred. They tested amino acid substitution within a conserved region of DME and found that the substitution was stable and heritable. They also investigated the status of methylation in the edited regions and found unaffected DNA methylation of the targeted genomic locus. These results were easily determined using PCR methods.
However, researchers concluded that further study must be done to better understand and improve gene targeting through HDR, so that it can be applied to other plants, especially crops.
For more information, read the article in Nature Communications.
The Crop Biotech Update is a weekly newsletter of ISAAA, a non-for-profit organization. The CBU is distributed for free to over 23,000 subscribers worldwide to inform them about the key developments in biosciences, especially in agricultural biotechnology. Your support will help us in our mission to feed the world with knowledge. You can help by donating as little as $10.
See more articles:
News from Around the World
- AfDB and FAO Boost Join Efforts to End Hunger and Create Wealth in Africa
- Purdue Researchers Discover Key to Mass Producing Beneficial Plant Compounds
- Researchers Unlock Secrets of Plant Development
- Argentina to Release First Biotech Soybean from China
- Farmers in South America Reap Significant Benefits from Planting Biotech Soybeans
- APHIS Announces Deregulation of GE Canola
- Researchers Discover Proteins Allow Plants to Rest Photosynthetic Activity at Night
- Japan's Ministry of Environment Meets to Discuss Genome Editing Regulation
- Scientists Isolate First Major Resistance Genes Against Wheat Stripe Rust Disease
- Global Biotech Crop Adoption Report Launched in Bogor, Indonesia
- Plant Leaves Possess A Molecular Compass
- Scientists Identify Maize QTLs for Simultaneous Root and Agronomic Trait Selection
Plant Breeding Innovations
- CRISPR-Cas9 Mediates Targeted Gene Replacement and Knockin in Arabidopsis
- Genome Editing to Control Gene Expression in Plants
- CRISPR-Cas9 Used to Reduce Symptoms of Cassava Brown Streak Disease
Beyond Crop Biotech
- Scientists Use Environmental DNA Approach for Agri Pest Surveillance
- Most Americans Accept GE Animals for Human Health, Study
- Guide to Understanding CRISPR Data Analysis Scores
Subscribe to CBU: