Our latest technology was published in Scientific Reports. This latest development will allow us to perform multiple transformation events.
Direct assembly of multiple linear DNA fragments via homologous recombination, a phenomenon known as in vivo assembly is used in biotechnology to assemble DNA constructs ranging in size from a few kilobases to full synthetic microbial genomes (see the figure for an illustration of how it works). Now this is also possible in the moss Physcomitrella patens.
It is an efficient method for gene deletion, gene insertion and total replacement of genes, genome parts and so on. Below is the integration of a fluorescents gene under the control of a endogenous promoter.