PubMed 33471325

PubMed ID: 33471325

View on PubMed
Agrobacterium tumefaciens-Mediated Transformation of Rice by Hygromycin Phosphotransferase (hptII) Gene Containing CRISPR/Cas9 Vector.
Authors: Majumder Shuvobrata, Datta Karabi, Datta Swapan Kumar
Journal: Methods in molecular biology (Clifton, N.J.) (Methods Mol Biol), Vol.2238(), 2021

DOI: 10.1111/j.1467-7652.2006.00196.x PMCID: 5554252

Abstract
The CRISPR/Cas9 technique for rice genome engineering is gaining momentum and requires a precise gene delivery system. For rice and other crop plants, Agrobacterium tumefaciens-mediated transformation (AMT) is considered a suitable gene transformation method. The AMT for indica-type rice is a challenge because it is less efficient in tissue culture response than japonica-type rice. Here is a protocol of the AMT method that we developed for IR64 variety which has been successfully tested in other popular indica-type rice varieties. We used embryogenic calli as explant and an empty gRNA-containing CRISPR/Cas9 vector with hptII (hygromycin phosphotransferase) gene for the transformation. This technique would speed up rice genome editing via CRISPR/Cas9 technology and facilitate to achieve varied application in the future.
Publication Types
Journal Article Research Support, Non-U.S. Gov't
Keywords
Agrobacterium-mediated transformation CRISPR/Cas9 Embryogenic calli IR64 Indica rice Rice Science Rice genome editing
Related Articles