A Multipurpose Toolkit to Enable Advanced Genome Engineering in Plants

The Plant Cell
Tomas CermakDaniel F Voytas

Abstract

We report a comprehensive toolkit that enables targeted, specific modification of monocot and dicot genomes using a variety of genome engineering approaches. Our reagents, based on transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system, are systematized for fast, modular cloning and accommodate diverse regulatory sequences to drive reagent expression. Vectors are optimized to create either single or multiple gene knockouts and large chromosomal deletions. Moreover, integration of geminivirus-based vectors enables precise gene editing through homologous recombination. Regulation of transcription is also possible. A Web-based tool streamlines vector selection and construction. One advantage of our platform is the use of the Csy-type (CRISPR system yersinia) ribonuclease 4 (Csy4) and tRNA processing enzymes to simultaneously express multiple guide RNAs (gRNAs). For example, we demonstrate targeted deletions in up to six genes by expressing 12 gRNAs from a single transcript. Csy4 and tRNA expression systems are almost twice as effective in inducing mutations as gRNAs expressed from individual RNA polymerase III promoters. Mutagenesis can be fur...Continue Reading

Citations

Mar 13, 2019·The Journal of Immunology : Official Journal of the American Association of Immunologists·Dmitri I KotovMarc K Jenkins
Jan 10, 2019·PloS One·Baptiste CastelJonathan D G Jones
Feb 23, 2019·Nucleic Acids Research·Ibtissam JabreNaeem H Syed
Mar 23, 2019·The Plant Journal : for Cell and Molecular Biology·Carla SchmidtHolger Puchta
May 1, 2019·Annual Review of Plant Biology·Erwang ChenBin Han
Oct 28, 2019·Horticulture Research·Jiemeng XuZhaobo Lang
Oct 17, 2017·Journal of Experimental Botany·Johann PetitChristophe Rothan
Mar 15, 2020·Plant Biotechnology Journal·Nathaniel M ButlerJiming Jiang
Jul 10, 2020·Journal of Experimental Botany·Rosa L López-MarquésMichael Palmgren
Jul 14, 2019·Scientific Reports·David ChiassonMartin Parniske
Jul 20, 2019·Transgenic Research·Caixia Gao
Jul 28, 2020·The Plant Journal : for Cell and Molecular Biology·Dae Kwan Ko, Federica Brandizzi
Aug 14, 2020·The Plant Journal : for Cell and Molecular Biology·Trevor WeissFeng Zhang
Jul 30, 2020·The Journal of Biological Chemistry·Joyce Van Eck
May 26, 2018·Plant Biotechnology Journal·Laurence TomlinsonJonathan D G Jones
Jul 19, 2018·Frontiers in Plant Science·Ryosuke HashimotoKeishi Osakabe
Nov 24, 2018·Journal of Integrative Plant Biology·Uwe Sonnewald
Mar 6, 2019·Annual Review of Plant Biology·Kunling ChenCaixia Gao
Feb 13, 2019·Journal of Experimental Botany·Grace L ChongloiUsha Vijayraghavan
Apr 21, 2018·Frontiers in Plant Science·Florian HahnAndreas P M Weber
Jun 10, 2018·G3 : Genes - Genomes - Genetics·Laurens PauwelsAlain Goossens
Oct 17, 2017·Frontiers in Plant Science·Chuanmei ZhuChristine Shyu
Apr 27, 2018·Scientific Reports·Pankaj BhowmikSateesh Kagale
Oct 3, 2018·Nature Biotechnology·Tingdong LiCao Xu
Apr 9, 2019·BioMed Research International·Nikolai BorisjukYuri Shavrukov
Dec 18, 2019·Nature Biotechnology·Michael F MaherDaniel F Voytas
Jun 3, 2020·Nature Plants·Evan E EllisonDaniel F Voytas

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