BROWSE

Related Scientist

Kang, Beum-Chang's photo.

Kang, Beum-Chang
유전체 교정 연구단
more info

ITEM VIEW & DOWNLOAD

Rapid delivery of Cas9 gene into the tomato cv. 'Heinz 1706' through an optimized Agrobacterium-mediated transformation procedure

Cited 0 time in webofscience Cited 0 time in scopus
17 Viewed 0 Downloaded
Title
Rapid delivery of Cas9 gene into the tomato cv. 'Heinz 1706' through an optimized Agrobacterium-mediated transformation procedure
Author(s)
BEEMNET MENGESHA KASSAHUN; BEUM-CHANG KANG; SU-JI BAE; YE JIN NAM; GRETEL FONSECA MUNDO; GA-HUI KANG; KYOUNGOOK KIM; JEUNG-SUL HAN
Publication Date
2021-01
Journal
Biocell, v.45, no.1, pp.199 - 215
Publisher
Tech Science Press
Abstract
© 2021 Centro Regional de Invest. Cientif. y Tecn.. All rights reserved.Solanum lycopersicum 'Heinz 1706' is a pioneer model cultivar for tomato research, whose whole genome sequence valuable for genomics studies is available. Nevertheless, a genetic transformation procedure for this cultivar has not yet been reported. Meanwhile, various genome editing technologies such as transfection of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) ribonucleoprotein complexes into cells are in the limelight. Utilizing the Cas9-expressing genotype possessing a reference genome can simplify the verification of an off-target effect, resolve the economic cost of Cas9 endonuclease preparation, and avoid the complex assembly process together with single-guide RNA (sgRNA) in the transfection approach. Thus, this study was designed to generate Cas9-expressing 'Heinz 1706' lines by establishing an Agrobacterium tumefaciens-mediated transformation (ATMT) procedure. Here, we report a rapid and reproducible transformation procedure for 'Heinz 1706' by finetuning various factors: A. tumefaciens strain, pre-culture and co-culture durations, a proper combination of phytohormones at each step, supplementation of acetosyringone, and shooting/rooting method. Particularly, through eluding subculture and simultaneously inducing shoot elongation and rooting from leaf cluster, we achieved a short duration of three months for recovering the transgenic plants expressing Cas9. The presence of the Cas9 gene and its stable expression were confirmed by PCR and qRT-PCR analyses, and the Cas9 gene integrated into the T0 plant genome was stably transmitted to T1 progeny. Therefore, we anticipate that our procedure appears to ease the conventional ATMT in 'Heinz 1706', and the created Cas9-expressing 'Heinz 1706' lines are ultimately useful in gene editing via unilateral transfection of sgRNA into the protoplasts.
URI
https://pr.ibs.re.kr/handle/8788114/9503
DOI
10.32604/BIOCELL.2021.012353
ISSN
0327-9545
Appears in Collections:
Center for Genome Engineering(유전체 교정 연구단) > 1. Journal Papers (저널논문)
Files in This Item:
There are no files associated with this item.

qrcode

  • facebook

    twitter

  • Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.
해당 아이템을 이메일로 공유하기 원하시면 인증을 거치시기 바랍니다.

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

Browse