CRISPR-transient expression in soybean for simplified gRNA screening in planta

Abstract The objective of this work was to develop a method create and validate CRISPR-Cas systems different gRNAs in soybean (Glycine max) embryos. Two model genes were used for simple mutation with one gRNA or partial gene deletion two guides. inserted into the CRISPR transformation vectors by type IIS restriction enzyme subcloning inserting promoter + gRNA2 final vector using classic cloning method. successfully constructed gRNAs. Agrobacterium-mediated transient carried out test quality system itself (expression cassette). Simple detected embryos transformed after DNA enrichment digestion followed polymerase chain reaction sequencing, which indicates that guides working. This protocol can be accelerate CRISPR-based genome editing strategies genetic soybean.