The Obstacles and Potential Solution Clues of Prime Editing Applications in Tomato

作     者：Tien Van Vu Ngan Thi Nguyen Jihae Kim Swati Das Jinsu Lee Jae-Yean Kim Tien Van Vu;Ngan Thi Nguyen;Jihae Kim;Swati Das;Jinsu Lee;Jae-Yean Kim

作者机构：Division of Applied Life Science(BK21 FOUR Program)Plant Molecular Biology and Biotechnology Research CenterGyeongsang National UniversityJinju 660-701Republic of Korea National Key Laboratory for Plant Cell BiotechnologyAgricultural Genetics InstituteKm 02Pham Van Dong RoadCo Nhue 1Bac Tu LiemHanoi 11917Vietnam Division of Life ScienceGyeongsang National University501 Jinju-daeroJinju 52828Republic of Korea Nulla Bio Inc.501 Jinju-daeroJinju 660-701Republic of Korea. 

基　　金：the National Research Foundation of Korea(the Bio&Medical Technology Development Program 2020M3A9I4038352,2020R1A6A1A03044344,2020R1I1A1A01072130,and 2022R1A2C3010331) the Pro gram for New Plant Breeding Techniques(NBT grant PJ016867022022) Rural Development Administration(RDA),Republic of Korea.Author contributions:T.V.V.and J.-Y.K.conceived and designed the research.T.V.V.,N.T.N.,J.K.,S.D.,and J.L.conducted the experiments.T.V.V.and J.-Y.K.analyzed the data.T.V.V.wrote the manuscript.T.V.V.and J.-Y.K.finalized the manuscript.All authors read and approved the manuscript 

出 版 物：《BioDesign Research》 (生物设计研究（英文）)

年 卷 期：2022年第2022卷第1期

页      码：11-18页

摘      要：Precision genome editing is highly desired for crop *** recently emerged CRISPR/Cas technology offers great potential applications in precision plant genome engineering.A prime editing(PE)approach combining a reverse transcriptase(RT)with a Cas9 nickase and a“priming”extended guide RNA(gRNA)has shown a high frequency for precise genome modification in mammalian cells and several plant ***,the applications of the PE approach in dicot plants are still limited and *** designed and tested prime editors for precision editing of a synthetic sequence in a transient assay and for desirable alleles of 10 loci in tomato by stable *** data obtained by targeted deep sequencing also revealed only low PE efficiencies in both the tobacco and tomato *** assessment of the activities of the PE components uncovered that the fusion of RT to Cas9 and the structure of PE gRNAs(pegRNAs)negatively affected the cleaving activity of the Cas9 *** self-complementarity between the primer binding sequences(PBSs)and spacer sequence might pose risks to the activity of the Cas9 ***,modifying the pegRNA sequences by shortening or introducing mismatches to the PBSs to reduce their melting temperatures did not enhance the PE efficiency at the MADS-box protein(SlMBP21),alcobaca(SlALC),and acetolactate synthase 1(SlALS1)*** data show challenges of the PE approach in tomato,indicating that a further improvement of the PE system for successful applications is demanded,such as the use of improved expression systems for enriching active PE complexes.

主 题 词：CRISPR/Cas precise editing 

学科分类：09[农学] 0902[农学-自然保护与环境生态类] 090202[090202] 

D　O　I：10.34133/bdr.0001

馆 藏 号：203121668...