| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | … The pRGEB32 vector used here was modified by replacing the hygromycin selectable marker with the BAR gene (confers glufosinate-ammonium resistance), facilitating this vector's use in maize?… Synthetic genes (from Genscript.com, capable of synthesizing highly?… | Get A Quote |
The CRISPR/Cas9-based system for targeted mutagenesis has become an indispensable tool for functional genetics in plants. CRISPR/Cas9 allows users to generate loss-of-function alleles in genes of interest with precision and in a simple-to-use system. This manuscript outlines important points to consider for experimental design and utilization of CRISPR/Cas9 in targeted mutagenesis in maize. It also introduces the pRGEB32-BAR vector modified for use in maize that allows simultaneous delivery of multiple gRNAs using a simple assembly. Vector selection, gRNA design, genetic strategies, and genotyping approaches are discussed, with an emphasis on achieving isolation of homozygous mutant plants in a time- and cost-e... More
