This chapter discusses the challenges facing woody perennial trees, namely the heterozygosity of outcrossing species and limited genomic resources. Gene knockouts (KO) represent the predominant applications of the CRISPR technology in woody perennial crops to date and will be the focus of this chapter. The issues considered for gene KO are generally applicable to other CRISPR applications targeting regulatory sequences or non-coding genes. Case studies from the Populus 4-coumarate:CoA ligase (4CL) family are presented to demonstrate the power of CRISPR in elucidating functional redundancy as well as specificity of duplicated genes in a perennial woody species.
Table of contents
2 The heterozygous genome challenge in gene editing
3 Genome resource limitation of woody perennial crops
4 Gene-editing reagents
6 CRISPR/Cas9-empowered characterization of duplicated genes
7 Conclusion and future trends
8 Where to look for further information