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Title: Diverse Systems for Efficient Sequence Insertion and Replacement in Precise Plant Genome Editing
CRISPR-mediated genome editing has been widely applied in plants to make uncomplicated genomic modifications including gene knockout and base changes. However, the introduction of many genetic variants related to valuable agronomic traits requires complex and precise DNA changes. Different CRISPR systems have been developed to achieve efficient sequence insertion and replacement but with limited success. A recent study has significantly improved NHEJ- and HDR-mediated sequence insertion and replacement using chemically modified donor templates. Together with other newly developed precise editing systems, such as prime editing and CRISPR-associated transposases, these technologies will provide new avenues to further the plant genome editing field.  more » « less
Award ID(s):
2029889 1758745
Author(s) / Creator(s):
Date Published:
Journal Name:
BioDesign Research
Page Range / eLocation ID:
1 to 4
Medium: X
Sponsoring Org:
National Science Foundation
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  2. Abstract

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  5. Abstract

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    Basic Protocol 1: Constructing the Cas9/sgRNA transient expression vector

    Alternate Protocol 1: Shortcut to generating single and pooled Cas9/sgRNA expression vectors

    Basic Protocol 2: Designing the oligonucleotide‐based homology‐directed repair (HDR) template

    Alternate Protocol 2: Designing the plasmid‐based HDR template

    Basic Protocol 3: Inducing genome editing by transforming CRISPR vector intoP. patensprotoplasts

    Basic Protocol 4: Identifying edited plants.

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