• Rapid production of SaCas9 in plant‐based cell‐free lysate for activity testing

    29 days ago - By Wiley

    Abstract
    Cas9 nucleases have become the most versatile tool for genome editing projects in a broad range of organisms. The recombinant production of Cas9 nuclease is desirable for in vitro activity assays or the preparation of ribonucleoproteins for DNA-free genome editing approaches. For the rapid production of Cas9, we explored the use of a recently established cell-free lysate from tobacco BY-2 cells. Using this system, the 130-kDa Cas9 nuclease from Staphylococcus aureus was produced and subsequently purified via affinity chromatography. The purified apoenzyme was supplemented with ten...
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  • Simplifying plant gene silencing and genome editing logistics by a one‐Agrobacterium system for simultaneous delivery of multipartite virus vectors

    Simplifying plant gene silencing and genome editing logistics by a one‐Agrobacterium system for simultaneous delivery of multipartite virus vectors

    29 days ago - By Wiley

    Graphical Abstract and Lay Summary
    Viral vectors are powerful tools for exogenous sequence delivery in plant cells. In this study, JoinTRV is designed for simultaneous delivery of engineered tobacco rattle virus RNA1 and RNA2 components from a single Agrobacterium cell in a 1-strain/2-vector system. JoinTRV allows flexible plant delivery of multi-component viral vectors as well as overexpression of genes of interest , virus-induced gene silencing and genome editing .
    Abstract
    Viral vectors provide a quick and effective way to express exogenous sequences in eukaryotic cells and to engineer...
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