18 January 2024

Heritable, multinucleotide deletions in plants using viral delivery of a repair exonuclease and guide RNAs

Degao Liu, Erik A Myers, Shuya Xuan, Lynn E Prichard, Lilee I Donahue, Evan E Ellison, Colby G Starker, Daniel F Voytas - Plant Physiology, 2024


CRISPR/Cas9-mediated mutagenesis typically results in short insertion/deletion mutations, which are often too small to disrupt the function of cis-acting regulatory elements. Here, we describe a highly efficient in planta gene editing approach called VirTREX2-HLDel that achieves heritable multinucleotide deletions in both protein-coding genes and noncoding DNA regulatory elements. VirTREX2-HLDel uses RNA viruses to deliver both the 3 prime repair exonuclease 2 (TREX2) and single-guide RNAs. Our method enables recovery of multiplexed heritable deletions and increases the heritable gene editing frequency at poorly edited sites. We identified functional conservation and divergence of MICRORNA164 (miR164) in Nicotiana benthamiana and tomato (Solanum lycopersicum) using VirTREX2-HLDel and observed previously uncharacterized phenotypes in plants with large deletions at this locus. Our viral delivery method reduces the need for tissue culture and will accelerate the understanding of protein-coding and regulatory regions in plants.

Read the entire publication

Share this:

Receive updates from the Crop Science Centre

Thanks! We’ve received your email.

Sorry, something went wrong.

By submitting this form, you are consenting to receive marketing emails from: The Crop Science Centre, Lawrence Weaver Rd, Cambridge, CB3 0LE, GB. You can revoke your consent to receive emails at any time by using the SafeUnsubscribe® link, found at the bottom of every email. Emails are serviced by Constant Contact.