TL;DR
This work compares protoplast transfection with emerging alternatives, including tissue electroporation and nanomaterial-, viral-, and mRNA-based delivery, and argues that progress will depend less on editing chemistry than on integrating delivery, tissue culture, regeneration, and validation into genotype-responsive workflows.
iPB-REG is established as a practical strategy for producing uniform genome-edited fruit trees and provide a valuable platform for DNA-free genetic improvement and functional genomics in clonally propagated perennial crops.
An efficient protocol for protoplast isolation from embryogenic calli and protoplast-to-plant regeneration in the allotetraploid species Coffea arabica is presented and polyethylene glycol-mediated transfection of coffee protoplasts with a CRISPR–Cas9 plasmid targeting the coffee xanthosine methyltransferase gene invol...
A critical review evaluates the strength and limits of the evidence for the claim that virus-derived products are transgene-free, and prioritised research directions are proposed, emphasising standardised reporting of progeny-level outcomes, biocontained vector design, and genotype-spanning validation in elite crop bac...
A stepwise protocol for targeted gene editing in wheat via wide hybridization with transgenic maize expressing Cas9 and gRNA is described, which provides a one-step approach for generating DH lines with the target gene edited from any wheat genotypes of interest.
An RNA-guided bridge recombinase system is engineered through rational mutagenesis and AI-assisted directed evolution, enabling programmable chromosomal rearrangements in both plant and mammalian cells and achieving up to a 29.8-fold increase in activity.
A novel plant genome editing system is developed by delivering Cas9, Cas12f, and Cas12j nucleases along with their guide RNAs using a broad-host-range geminivirus, Wheat dwarf India virus (WDIV), in combination with Ageratum yellow leaf curl betasatellite (AYLCB).
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