Heritable CRISPR-Cas9 editing of plant genomes using RNA virus vectors

Viral vectors hold enormous potential for genome editing in plants as transient delivery vehicles of CRISPR-Cas components. Here, we describe a protocol to assemble plant viral vectors for single-guide RNA (sgRNA) delivery. The obtained viral constructs are based on compact T-DNA binary vectors of the pLX series and are delivered into Cas9-expressing plants through agroinoculation. This approach allows rapidly assessing sgRNA design for plant genome targeting, as well as the recovery of progeny with heritable mutations at targeted loci. For complete details on the use and execution of this protocol, please refer to Uranga et al. (2021)1 and Aragone ' s et al. (2022).2

Automated summary

Viral vectors are powerful tools for editing plant genomes. This protocol describes a method for assembling plant viral vectors for sgRNA delivery, allowing rapid assessment of design effectiveness and the generation of heritable mutations.