Tissue Culture and Genome Editing
Project Overview:
Plant transformation has become an integral part of plant genetics over the last few decades. For most of this time, researchers have relied on random integrations of novel DNA sequences that cause either a gain of function or loss of function of a gene. New technologies such as CRISPR-Cas9 now enable the precise modification of genes within the maize genome.
The Moose Lab is currently developing a system that utilizes this technology in conjunction with micropropagation and biolistic transformation in the Illinois Long Term Selection Experiment. We have developed a reliable tissue culture protocol for regenerating the Illinois High Protein (IHP) and Illinois Low Protein (ILP) genotypes. Our next goal is to integrate a single transcriptional unit (STU) construct that contains Cas9 and specific guide RNAs to edit genes of interest identified from the Long Term Selection Experiment. We expect this system to be a robust tool for helping us dissect the genetic basis of the divergence of IHP and ILP and use this information to validate other experiments pertaining to nitrogen use and protein accumulation.
An overview of the induction, transformation, and regeneration of the IHP and ILP genotypes in tissue culture. Both genotypes produce distinct morphological characteristics of callus growth and exhibit different regeneration frequencies.