An improved biolistic delivery and analysis method for evaluation of DNA and CRISPR-Cas delivery efficacy in plant tissue

Abstract Biolistic delivery is widely used for genetic transformation but inconsistency between bombardment samples transient gene expression analysis often hinders quantitative analyses. We developed a methodology to improve the consistency of biolistic results by using double-barrel device and cell counting software. The enables strategy incorporating an internal control into each sample, which significantly decreases variance results. software further reduces errors increases throughput. utility this new platform demonstrated optimizing conditions delivering DNA commercial transfection reagent Trans IT-2020. In addition, same approach applied test efficacy multiple gRNAs CRISPR-Cas9-mediated editing. novel combination method allows simultaneous comparison optimization parameters in delivery. here can be broadly any target biolistics, including animal cells tissues.