CRISPR Milestones for Sustainable Agriculture and Forestry

Summary: The CRISPR-Cas9 system was used to perform precise mutagenesis in a tandem array of seven genes in Populus tremula x Populus alba. Efficient multiplex editing was achieved with a single guide RNA in 42 transgenic lines, resulting in a range of mutation profiles including small insertions/deletions, local deletions, large genomic rearrangements, and complex rearrangements. Target capture sequencing was used to analyze repair outcomes and reconstruct mutant alleles. This work demonstrates the power of CRISPR-Cas9 for generating diverse mutants with structural and copy number variations in tandemly duplicated genes.