AI Article Synopsis
- Viral vectors are being explored as effective tools for delivering CRISPR-Cas components to edit plant genomes.
- The protocol involves assembling viral vectors specifically for delivering single-guide RNA (sgRNA) using compact T-DNA binary vectors, which are then introduced into Cas9-expressing plants through a technique called agroinoculation.
- This method facilitates rapid evaluation of sgRNA designs and enables the recovery of offspring with heritable mutations in targeted genetic locations.
Article Abstract
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) and Aragonés et al. (2022)..
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9943877 | PMC |
| http://dx.doi.org/10.1016/j.xpro.2023.102091 | DOI Listing |
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