Li Gen, Zhang Yingxiao, Dailey Micah, Qi Yiping
Department of Plant Science and Landscape Architecture, University of Maryland, College Park, College Park, MD, United States.
Institute for Bioscience and Biotechnology Research, University of Maryland, Rockville, MD, United States.
Front Genome Ed. 2023 Oct 12;5:1251903. doi: 10.3389/fgeed.2023.1251903. eCollection 2023.
Cas12a, also known as Cpf1, is a highly versatile CRISPR-Cas enzyme that has been widely used in genome editing. Unlike its well-known counterpart, Cas9, Cas12a has unique features that make it a highly efficient genome editing tool at AT-rich genomic regions. To enrich the CRISPR-Cas12a plant genome editing toolbox, we explored 17 novel Cas12a orthologs for their genome editing capabilities in plants. Out of them, Ev1Cas12a and Hs1Cas12a showed efficient multiplexed genome editing in rice and tomato protoplasts. Notably, Hs1Cas12a exhibited greater tolerance to lower temperatures. Moreover, Hs1Cas12a generated up to 87.5% biallelic editing in rice T plants. Both Ev1Cas12a and Hs1Cas12a achieved effective editing in poplar T plants, with up to 100% of plants edited, albeit with high chimerism. Taken together, the efficient genome editing demonstrated by Ev1Cas12a and Hs1Cas12a in both monocot and dicot plants highlights their potential as promising genome editing tools in plant species and beyond.
Cas12a，也被称为Cpf1，是一种用途广泛的CRISPR-Cas酶，已被广泛应用于基因组编辑。与广为人知的Cas9不同，Cas12a具有独特的特性，使其成为富含AT的基因组区域的高效基因组编辑工具。为了丰富CRISPR-Cas12a植物基因组编辑工具箱，我们探索了17种新型Cas12a直系同源物在植物中的基因组编辑能力。其中，Ev1Cas12a和Hs1Cas12a在水稻和番茄原生质体中表现出高效的多重基因组编辑。值得注意的是，Hs1Cas12a对较低温度表现出更大的耐受性。此外，Hs1Cas12a在水稻T0植株中产生了高达87.5%的双等位基因编辑。Ev1Cas12a和Hs1Cas12a在杨树T0植株中均实现了有效的编辑，尽管嵌合体比例较高，但高达100%的植株被编辑。综上所述，Ev1Cas12a和Hs1Cas12a在单子叶植物和双子叶植物中均表现出高效的基因组编辑，突出了它们作为植物及其他物种中有前景的基因组编辑工具的潜力。
