Plant Science

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    Protocols in Current Issue
    Construction of Antisense RNA-mediated Gene Knock-down Strains in the Cyanobacterium Anabaena sp. PCC 7120
    Authors:  Amit Srivastava, Anand Ballal, Karl Forchhammer and Anil Kumar Tripathi, date: 02/20/2020, view: 948, Q&A: 0
    [Abstract] Anabaena sp. PCC 7120 (hereafter Anabaena) is a model cyanobacterium to study nitrogen fixation, cellular differentiation and several other key biological functions that are analogous in plants. As with any other organism, many genes in Anabaena encode an essential life function and hence cannot be deleted, causing a ...
    High Resolution Melting Temperature Analysis to Identify CRISPR/Cas9 Mutants from Arabidopsis
    Authors:  Cynthia Denbow, Sonia Carole Ehivet and Sakiko Okumoto, date: 07/20/2018, view: 4123, Q&A: 0
    [Abstract] CRISPR/Cas9 made targeted mutagenesis and genome editing possible for many plant species. One of the ways that the endonuclease is used for plant genetics is the creation of loss-of-function mutants, which typically result from erroneous DNA repair through non-homologous end joining (NHEJ) pathway. The majority of erroneous repair events results ...
    Identification of Insertion Site by RESDA-PCR in Chlamydomonas Mutants Generated by AphVIII Random Insertional Mutagenesis
    Authors:  Fantao Kong and Yonghua Li-Beisson, date: 02/05/2018, view: 4044, Q&A: 0
    [Abstract] Chlamydomonas reinhardtii is frequently used as a model organism to study fundamental processes in photosynthesis, metabolism, and flagellar biology. Versatile tool boxes have been developed for this alga (Fuhrmann et al., 1999; Schroda et al., 2000; Schroda, 2006). Among them, forward genetic approach has been ...
    Construction of a Single Transcriptional Unit for Expression of Cas9 and Single-guide RNAs for Genome Editing in Plants
    Authors:  Xu Tang, Zhaohui Zhong, Xuelian Zheng and Yong Zhang, date: 09/05/2017, view: 7907, Q&A: 0
    [Abstract] The CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein9 (Cas9) is a simple and efficient tool for genome editing in many organisms including plant and crop species. The sgRNAs of the CRISPR/Cas9 system are typically expressed from RNA polymerase III promoters, such as U6 and U3. In many transformation events, ...
    Generation of Targeted Knockout Mutants in Arabidopsis thaliana Using CRISPR/Cas9
    Authors:  Florian Hahn, Marion Eisenhut, Otho Mantegazza and Andreas P. M. Weber, date: 07/05/2017, view: 14676, Q&A: 1
    [Abstract] The CRISPR/Cas9 system has emerged as a powerful tool for gene editing in plants and beyond. We have developed a plant vector system for targeted Cas9-dependent mutagenesis of genes in up to two different target sites in Arabidopsis thaliana. This protocol describes a simple 1-week cloning procedure for a single T-DNA vector containing ...
    Targeted Mutagenesis Using RNA-guided Endonucleases in Mosses
    Authors:  Toshihisa Nomura and Hitoshi Sakakibara, date: 06/20/2017, view: 7532, Q&A: 0
    [Abstract] RNA-guided endonucleases (RGENs) have been used for genome editing in various organisms. Here, we demonstrate a simple method for performing targeted mutagenesis and genotyping in a model moss species, Physcomitrella patens, using RGENs. We also performed targeted mutagenesis in a non-model moss, Scopelophilla cataractae, using a ...
    Strategies for Performing Dynamic Gene Perturbation Experiments in Flowers
    Authors:  Diarmuid S. Ó’Maoiléidigh, Emmanuelle Graciet and Frank Wellmer, date: 04/05/2016, view: 5627, Q&A: 0
    [Abstract] Dissecting the gene regulatory networks (GRNs) underlying developmental processes is a central goal in biology. The characterization of the GRNs underlying flower development has received considerable attention, however, novel approaches are required to reveal temporal and spatial aspects of these GRNs. Here, we provide an overview of the options ...



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