# T-DNAreader: fast and precise identification of T-DNA insertion sites in plant genomes using RNA sequencing data

**Authors:** Hongwoo Lee, Pil Joon Seo

PMC · DOI: 10.1186/s13059-025-03655-x · 2025-07-10

## TL;DR

T-DNAreader is a fast and accurate tool for identifying where T-DNA inserts into plant genomes using RNA sequencing data.

## Contribution

T-DNAreader improves precision, sensitivity, and speed in identifying T-DNA insertion sites compared to existing tools.

## Key findings

- T-DNAreader efficiently identifies T-DNA insertion sites within transcribed regions.
- The tool outperforms existing methods in precision and speed.
- It standardizes the characterization of T-DNA-containing transgenic plants.

## Abstract

Agrobacterium-mediated plant transformation, which enables the delivery of DNA using transfer DNA (T-DNA) binary vectors, is an essential technique in plant research. T-DNAs randomly integrate into the host genome, and multiple T-DNAs can sometimes integrate during a single transformation, necessitating the development of tools to elucidate the T-DNA insertion sites (TISs). Here, we present T-DNAreader, which identifies TISs from RNA sequencing data with high precision, sensitivity, and speed, outperforming existing tools. T-DNAreader enables the efficient and reliable identification of TISs within transcribed regions and standardizes the characterization of T-DNA-containing transgenic plants, which can be expanded to various organisms containing T-DNA.

The online version contains supplementary material available at 10.1186/s13059-025-03655-x.

## Full-text entities

- **Genes:** AT1G09807 (ncRNA) [NCBI Gene 28717175], RDR1 (RNA-dependent RNA polymerase 1) [NCBI Gene 838044] {aka ATRDRP1, AtRDR1, F10B6.19, F10B6_19, RNA-dependent RNA polymerase 1}, SUT2 (sucrose transporter 2) [NCBI Gene 814817] {aka ARABIDOPSIS THALIANA SUCROSE TRANSPORTER 3, ATSUC3, ATSUT2, SUC3, SUCROSE TRANSPORTER 3, T17M13.3}, AT4G09630 (transmembrane protein (DUF616)) [NCBI Gene 826549] {aka T25P22.70, T25P22_70}, AT3G26100 (Regulator of chromosome condensation (RCC1) family protein) [NCBI Gene 822208], ROPGEF3 (RHO guanyl-nucleotide exchange factor 3) [NCBI Gene 827967] {aka ATROPGEF3, A_IG005I10.1, A_IG005I10_1, KINASE PARTNER PROTEIN-LIKE, KPP-LIKE, RHO guanyl-nucleotide exchange factor 3}, PLT1 (Integrase-type DNA-binding superfamily protein) [NCBI Gene 821632] {aka PLETHORA 1}, FHY1 (far-red elongated hypocotyl 1) [NCBI Gene 5007942] {aka FAR-RED ELONGATED HYPOCOTYL 1, FRY1, PAT3, PHYTOCHROME A SIGNAL TRANSDUCTION 3, far-red elongated hypocotyl 1}, EXO70H4 (exocyst subunit exo70 family protein H4) [NCBI Gene 820109] {aka ATEXO70H4, exocyst subunit exo70 family protein H4}, PLT2 (Integrase-type DNA-binding superfamily protein) [NCBI Gene 841542] {aka F11M15.6, F11M15_6, PLETHORA 2}, AT5G12440 (CCCH-type zinc fingerfamily protein with RNA-binding domain-containing protein) [NCBI Gene 831119], NLP7 (NIN like protein 7) [NCBI Gene 828502] {aka NIN like protein 7, T19F6.10, T19F6_10}
- **Diseases:** TISs (MESH:D009371), crown gall disease (MESH:D005705)
- **Chemicals:** T-LOC (-)
- **Species:** Verticillium dahliae (species) [taxon 27337], Marchantia polymorpha (common liverwort, species) [taxon 3197], Hapsidospora chrysogena (species) [taxon 5044], Agrobacterium tumefaciens (species) [taxon 358], Zymoseptoria tritici (species) [taxon 1047171], Oryza sativa (Asian cultivated rice, species) [taxon 4530], Zea mays (maize, species) [taxon 4577], Arabidopsis thaliana (mouse-ear cress, species) [taxon 3702], Solanum lycopersicum (tomato, species) [taxon 4081]

## Figures

9 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12243177/full.md

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Source: https://tomesphere.com/paper/PMC12243177