Abstract
Genome editing is widely used across plant species to generate and study the impact of functional mutations in crop improvement. However, transgene integration in plant genomes raises important legislative concerns regarding genetically modified organisms. Several strategies have been developed to remove or prevent the integration of gene editor constructs, which can be divided into three major categories: 1) elimination of transgenic sequences via genetic segregation; 2) transient editor expression from DNA vectors; and 3) DNA-independent editor delivery, including RNA or preassembled Cas9 protein-gRNA ribonucleoproteins (RNPs). Here, we summarize the main strategies employed to date and discuss the advantages and disadvantages of using these different tools. We hope that our work can provide important information concerning the value of alternative genome editing strategies to advance crop breeding.
Highlights
Genome editing is a revolutionary technology for the advancement of plant science and crop breeding (Chen et al, 2019)
double-strand break (DSB) repair in plants is majorly achieved through an error prone non-homologous end joining (NHEJ) pathway, which usually leads to some base insertions/deletions and generates mutations at the target site (Jiang and Doudna, 2017)
clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system-mediated genome editing leads to efficient target modification in plants, including the model plant Arabidopsis and several crop species (Chen et al, 2019; Kong et al, 2021)
Summary
Received: 30 October 2021 Accepted: 12 November 2021 Published: 02 December 2021. Genome editing is widely used across plant species to generate and study the impact of functional mutations in crop improvement. Transgene integration in plant genomes raises important legislative concerns regarding genetically modified organisms. Several strategies have been developed to remove or prevent the integration of gene editor constructs, which can be divided into three major categories: 1) elimination of transgenic sequences via genetic segregation; 2) transient editor expression from DNA vectors; and 3) DNA-independent editor delivery, including RNA or preassembled Cas protein-gRNA ribonucleoproteins (RNPs). We summarize the main strategies employed to date and discuss the advantages and disadvantages of using these different tools. We hope that our work can provide important information concerning the value of alternative genome editing strategies to advance crop breeding
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