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??? 將CRISPR-Cas9蛋白和gRNA在體外組裝成核糖核蛋白復(fù)合體，再利用基因槍轉(zhuǎn)化，就可以得到全程無(wú)DNA作為載體的基因編輯作物產(chǎn)品，該產(chǎn)品的優(yōu)勢(shì)是編輯效率高、脫靶率低、非預(yù)期變異少、背景純合易。如今，種業(yè)巨頭杜邦先鋒公司已成功利用該技術(shù)得到了玉米基因編輯植株，性狀表現(xiàn)為磺酰脲類(lèi)除草劑抗性、雄性不育和葉舌缺失。

??? 繼雜交糯玉米之后，杜邦先鋒公司在玉米基因編輯技術(shù)上又有了新的斬獲，這是否意味著杜邦先鋒在作物編輯育種領(lǐng)域已經(jīng)領(lǐng)先了半個(gè)身位？

Nature Communications?7:?13274. 16 November 2016

Genome editing in maize directed by CRISPR–Cas9 ribonucleoprotein complexes

Author

Sergei Svitashev,?Christine Schwartz,?Brian Lenderts,?Joshua K. Young&?A. Mark Cigan

Trait Enabling Technologies, DuPont Pioneer, USA

Abstract

Targeted DNA double-strand breaks have been shown to significantly increase the frequency and precision of genome editing. In the past two decades, several double-strand break technologies have been developed. CRISPR–Cas9 has quickly become the technology of choice for genome editing due to its simplicity, efficiency and versatility. Currently, genome editing in plants primarily relies on delivering double-strand break reagents in the form of DNA vectors. Here we report biolistic delivery of pre-assembled Cas9–gRNA ribonucleoproteins into maize embryo cells and regeneration of plants with both mutated and edited alleles. Using this method of delivery, we also demonstrate DNA- and selectable marker-free gene mutagenesis in maize and recovery of plants with mutated alleles at high frequencies. These results open new opportunities to accelerate breeding practices in a wide variety of crop species.