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Protein expression and gene editing in monocots using foxtail mosaic virus vectors.

Plant Direct. 2019; 
Mei Y, Beernink BM, Ellison EE, Kone?ná E, Neelakandan AK, Voytas DF, Whitham SA.
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Monoclonal Antibody Services … After centrifugation, the supernatant was mixed with SDS‐PAGE loading buffer and boiled for 5 min, and then, 10 μl of each boiled sample was used in SDS‐PAGE followed by Western blotting using anti‐GFP monoclonal antibody (Genscript) … Get A Quote

摘要

Plant viruses can be engineered to carry sequences that direct silencing of target host genes, expression of heterologous proteins, or editing of host genes. A set of foxtail mosaic virus (FoMV) vectors was developed that can be used for transient gene expression and single guide RNA delivery for Cas9-mediated gene editing in maize, Setaria viridis, and Nicotiana benthamiana. This was accomplished by duplicating the FoMV capsid protein subgenomic promoter, abolishing the unnecessary open reading frame 5A, and inserting a cloning site containing unique restriction endonuclease cleavage sites immediately after the duplicated promoter. The modified FoMV vectors transiently expressed green fluorescent protein (GFP)... More

關鍵詞

CRISPR; Cas9; gene editing; gene expression; guide RNA; virus
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