# Genome Editing by Grafting

**Authors:** Samuel Simoni, Marco Fambrini, Claudio Pugliesi, Ugo Rogo

PMC · DOI: 10.3390/ijms26199294 · 2025-09-23

## TL;DR

This review explores how grafting can be used to edit plant genomes without introducing foreign DNA, offering a promising new method for crop improvement.

## Contribution

The paper introduces grafting as a novel, transgene-free method for delivering CRISPR components to edit plant genomes.

## Key findings

- Grafting allows delivery of CRISPR components via RNA without integrating foreign DNA into the genome.
- This method can be used for plants that are difficult to edit using traditional in vitro techniques.
- The technique shows potential for improving crop breeding but faces challenges like variable efficiency and graft incompatibility.

## Abstract

Grafting is the process of joining parts of two plants, allowing the exchange of molecules such as small RNAs (including microRNAs and small interfering RNAs), messenger RNAs, and proteins between the rootstock and the scion. Genome editing by grafting exploits RNAs, such as tRNA-like sequences (TLS motifs), to deliver the components (RNA) of the clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) system from transgenic rootstock to wild-type scion. The complex Cas9 protein and sgRNA-TLS produced in the scion perform the desired modification without the integration of foreign DNA in the plant genome, resulting in heritable transgene-free genome editing. In this review, we examine the current state of the art of this innovation and how it helps address regulatory problems, improves crop recovery and selection, exceeds the usage of viral vectors, and may reduce potential off-target effects. We also discuss the promise of genome editing by grafting for plants recalcitrant to in vitro culture and for agamic-propagated species that must maintain heterozygosity for plant productivity, fruit quality, and adaptation. Furthermore, we explore the limitations of this technique, including variable efficiency, graft incompatibility among genotypes, and challenges in large-scale application, while highlighting its considerable potential for further improvement and future broader applications for crop breeding.

## Linked entities

- **Proteins:** cas9 (type II CRISPR RNA-guided endonuclease Cas9)

## Full-text entities

- **Genes:** GAI (GRAS family transcription factor family protein) [NCBI Gene 838057] {aka F10B6.34, F10B6_34, GAI PROTEIN, GIBBERELLIC ACID INSENSITIVE, RESTORATION ON GROWTH ON AMMONIA 2, RGA2}, DMC1 (DNA repair (Rad51) family protein) [NCBI Gene 821860] {aka ARABIDOPSIS HOMOLOG OF LILY MESSAGES INDUCED AT MEIOSIS 15, ARABIDOPSIS THALIANA DISRUPTION OF MEIOTIC CONTROL 1, ARLIM15, ATDMC1, DISRUPTION OF MEIOTIC CONTROL 1}, WUS (Homeodomain-like superfamily protein) [NCBI Gene 816305] {aka PGA6, T27K22.18, T27K22_18, WUS1, WUSCHEL, WUSCHEL 1}, tRNAMet [NCBI Gene 4024959], NIA1 (nitrate reductase 1) [NCBI Gene 844112] {aka GNR1, NITRATE REDUCTASE, NITRATE REDUCTASE 1, NR1, T32E8.9, T32E8_9}, FT (PEBP (phosphatidylethanolamine-binding protein) family protein) [NCBI Gene 842859] {aka F5I14.3, F5I14_3, FLOWERING LOCUS T, REDUCED STEM BRANCHING 8, RSB8}, STM (KNOX/ELK homeobox transcription factor) [NCBI Gene 842534] {aka BUM, BUM1, BUMBERSHOOT, BUMBERSHOOT 1, F24O1.38, F24O1_38}, HSC70-1 (heat shock cognate protein 70-1) [NCBI Gene 831020] {aka ARABIDOPSIS THALIANA HEAT SHOCK COGNATE PROTEIN 70-1, AT-HSC70-1, AtHsp70-1, HEAT SHOCK COGNATE PROTEIN 70, HEAT SHOCK PROTEIN 70-1, HSC70}
- **Diseases:** water deficiency (MESH:D003681), toxicity (MESH:D064420), infection (MESH:D007239), GE (MESH:D042822), drought (MESH:C536747), TLS (OMIM:615281), injury to (MESH:D014947), Panama disease (MESH:D004194), late blight (MESH:D000067562), male sterility (MESH:D007248)
- **Chemicals:** starch (MESH:D013213), Fanzor (-), auxin (MESH:D007210), poly(A) (MESH:D011061), lipids (MESH:D008055), silicon carbide (MESH:C022088), cytokinins (MESH:D003583), carbon (MESH:D002244), PEG (MESH:D011092), nitrate (MESH:D009566)
- **Species:** Homo sapiens (human, species) [taxon 9606], TSWV [taxon 1933298], Brassica rapa (field mustard, species) [taxon 3711], Aequorea victoria (species) [taxon 6100], Arabidopsis thaliana (mouse-ear cress, species) [taxon 3702], Manihot esculenta (cassava, species) [taxon 3983], Solanum lycopersicum (tomato, species) [taxon 4081], Cytorhabdovirus hordei (no rank) [taxon 1985699], Persea americana (avocado, species) [taxon 3435], Triticum aestivum (bread wheat, species) [taxon 4565], Foxtail mosaic virus (no rank) [taxon 12179], Nicotiana benthamiana (species) [taxon 4100], Oryza sativa (Asian cultivated rice, species) [taxon 4530], Solanum tuberosum (potatoes, species) [taxon 4113], Sorghum bicolor (broomcorn, species) [taxon 4558], Theobroma cacao (cacao, species) [taxon 3641], Barley stripe mosaic virus (no rank) [taxon 12327], Turnip yellow mosaic virus (no rank) [taxon 12154], Musa acuminata AAA Group (Cavendish banana, genotype) [taxon 214697], Helianthus annuus (common sunflower, species) [taxon 4232], Prunus persica (peach, species) [taxon 3760], Capsicum (peppers, genus) [taxon 4071], Malus domestica (apple, species) [taxon 3750], Tobacco ringspot virus (no rank) [taxon 12282], Solanum pennellii (species) [taxon 28526], Cenchrus americanus (bulrush millet, species) [taxon 4543], Vitis vinifera (wine grape, species) [taxon 29760], Thanatephorus sp. RV (species) [taxon 359004], Potato virus X (no rank) [taxon 12183]

## Figures

2 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12524868/full.md

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Source: https://tomesphere.com/paper/PMC12524868