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    Home > Biochemistry News > Biotechnology News > The latest paper of Huazhong Agricultural University: A new high-throughput gene editing technology

    The latest paper of Huazhong Agricultural University: A new high-throughput gene editing technology

    • Last Update: 2021-10-21
    • Source: Internet
    • Author: User
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    Nanhu News Network (correspondent Li Shumin) Recently, Kabin Xie's group at the State Key Laboratory of Crop Genetic Improvement of Huazhong Agricultural University established an array-type CRISPR library for large-scale plant gene editing
    .


    Using this method, the research group created a gene editing material targeted to knock out 1072 rice receptor kinases, which provided new resources for the rapid identification of genes related to disease resistance and stress resistance


    With the increasing maturity of CRISPR/Cas9 gene editing technology in plants, it is possible to use CRISPR libraries for high-throughput genetic screening
    .


    There are currently two CRISPR library construction strategies: pooled library and arrayed library


    In this study, a rapid construction technology of arrayed CRISPR library was established, which can be used for both small-scale and large-scale plant gene editing materials creation
    .


    The author designed a strategy of PCR fragment-length markers for gRNAs distinguishing (FLASH tag for short) that uses PCR fragment-length markers for gRNAs distinguishing.


    The process of constructing a plant mutant library using the FLASH tag vector system

    Using the FLASH strategy, this research completed the construction of targeted editing 1072 rice receptor kinases in about a year
    .


    The research team used 12 vectors to transform rice and obtained 5,039 T0 rice plants through 89 rice transformation events.


    Targeted editing of rice RLK gene family

    In this study, off-target editing and gene editing events without T-DNA integration were also analyzed in detail, and the rice blast inoculation test was carried out on 15 gene materials, and 9 genes related to rice blast resistance were identified
    .


    Finally, the research group discussed the advantages and disadvantages of this method, and proposed a collaborative scheme for constructing a genome-wide array CRISPR library


    Collaborative solution for expanding array CRISPR library using FLASH gene editing pipeline

    The 2021 PhD graduate Chen Kaiyuan of our school is the first author of the paper, and Professor Xie Kabin is the corresponding author; Professor Xin Hou of Wuhan University, Professor Yang Yinong of Pennsylvania State University and Professor Xiong Lizhong of our school also participated in the research
    .


    The research was funded by the National Natural Science Foundation of China, the Major Project of Genetically Modification of the Ministry of Science and Technology, and the Independent Innovation Fund of Huazhong Agricultural University


    【English summary】

    Clustered regularly interspaced short palindromic repeats (CRISPR)−CRISPR-associated protein 9 (Cas9)-mediated gene editing is revolutionizing plant research and crop breeding.


    Link to the paper : https://doi.


    Reviewer: Xie Kabin

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