A5049418431- CRISPR and Genetic Engineering
- Plant Virus Research Studies
- Chromosomal and Genetic Variations
- Plant tissue culture and regeneration
- RNA and protein synthesis mechanisms
- Transgenic Plants and Applications
- Advanced biosensing and bioanalysis techniques
- Plant Disease Resistance and Genetics
- Prostate Cancer Treatment and Research
- RNA Research and Splicing
- Photosynthetic Processes and Mechanisms
- Fungal and yeast genetics research
- RNA Interference and Gene Delivery
- Insect symbiosis and bacterial influences
- Plant Molecular Biology Research
- RNA modifications and cancer
- Genomics and Phylogenetic Studies
- Legume Nitrogen Fixing Symbiosis
- Plant nutrient uptake and metabolism
- Plant Pathogenic Bacteria Studies
- Genomics and Chromatin Dynamics
- Microbial Community Ecology and Physiology
- Pluripotent Stem Cells Research
- Virus-based gene therapy research
- Animal Genetics and Reproduction
University of Minnesota
2016-2025
University of Minnesota System
2010-2025
Biotechnology Institute
2022-2024
Cargill (United States)
2013-2022
Institut thématique Génétique, génomique et bioinformatique
2007-2021
ORCID
2020
Donald Danforth Plant Science Center
2018
Charles University
2017
Czech Academy of Sciences, Institute of Physiology
2017
Czech Academy of Sciences, Institute of Molecular Genetics
2017
TALENs are important new tools for genome engineering. Fusions of transcription activator-like (TAL) effectors plant pathogenic Xanthomonas spp. to the FokI nuclease, bind and cleave DNA in pairs. Binding specificity is determined by customizable arrays polymorphic amino acid repeats TAL effectors. We present a method reagents efficiently assembling TALEN constructs with custom repeat arrays. also describe design guidelines based on naturally occurring their binding sites. Using software...
Engineered nucleases that cleave specific DNA sequences in vivo are valuable reagents for targeted mutagenesis. Here we report a new class of sequence-specific created by fusing transcription activator-like effectors (TALEs) to the catalytic domain FokI endonuclease. Both native and custom TALE-nuclease fusions direct double-strand breaks specific, sites.
The relative ease, speed, and biological scope of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated Protein9 (Cas9)-based reagents for genomic manipulations are revolutionizing virtually all areas molecular biosciences, including functional genomics, genetics, applied biomedical research, agricultural biotechnology. In plant systems, however, a number hurdles currently exist that limit this technology from reaching its full potential. For example,...
The use of homologous recombination to precisely modify plant genomes has been challenging, due the lack efficient methods for delivering DNA repair templates cells. Even with advent sequence-specific nucleases, which stimulate at predefined genomic sites by creating targeted double-strand breaks, there are only a handful studies that report precise editing endogenous genes in crop plants. More needed through recombination, ideally without randomly integrating foreign DNA. Here, we...
Transcription activator-like effector nucleases (TALENs) are programmable that join FokI endonuclease with the modular DNA-binding domain of TALEs. Although zinc-finger enable a variety genome modifications, their application to genetic engineering livestock has been slowed by technical limitations embryo-injection, culture primary cells, and difficulty in producing reliable reagents limited budget. In contrast, we found TALENs could easily be manufactured over half (23/36, 64%) demonstrate...
Transcription activator-like (TAL) effectors are repeat-containing proteins used by plant pathogenic bacteria to manipulate host gene expression. Repeats polymorphic and individually specify single nucleotides in the DNA target, with some degeneracy. A TAL effector-nucleotide binding code that links repeat type specified nucleotide enables prediction of genomic sites for customization use targeting, particular as custom transcription factors engineered regulation site-specific nucleases...
We report a comprehensive toolkit that enables targeted, specific modification of monocot and dicot genomes using variety genome engineering approaches. Our reagents, based on transcription activator-like effector nucleases (TALENs) the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system, are systematized for fast, modular cloning accommodate diverse regulatory sequences to drive reagent expression. Vectors optimized create either single or multiple gene knockouts...
We conducted a genome-wide survey of Saccharomyces cerevisiae retrotransposons and identified total 331 insertions, including 217 Ty1, 34 Ty2, 41 Ty3, 32 Ty4, 7 Ty5 elements. Eighty-five percent insertions were solo long terminal repeats (LTRs) or LTR fragments. Overall, retrotransposon sequences constitute >377 kb 3.1% the genome. Independent evolution was evidenced by identification single-base pair insertion/deletion that distinguishes highly similar Ty1 Ty2 LTRs distinct subfamily...
Summary Soybean oil is high in polyunsaturated fats and often partially hydrogenated to increase its shelf life improve oxidative stability. The trans‐fatty acids produced through hydrogenation pose a health threat. lines that are low were generated by introducing mutations two fatty acid desaturase 2 genes ( FAD 2‐1A 2‐1B ), which the seed convert monounsaturated fat, oleic acid, linoleic acid. Transcription activator‐like effector nucleases TALEN s) engineered recognize cleave conserved...
Coeliac disease is an autoimmune disorder triggered in genetically predisposed individuals by the ingestion of gluten proteins from wheat, barley and rye. The α-gliadin gene family wheat contains four highly stimulatory peptides, which 33-mer main immunodominant peptide patients with coeliac. We designed two sgRNAs to target a conserved region adjacent coding sequence for genes. Twenty-one mutant lines were generated, all showing strong reduction α-gliadins. Up 35 different genes mutated one...
Sequence-specific nucleases enable facile editing of higher eukaryotic genomic DNA; however, targeted modification plant genomes remains challenging due to ineffective methods for delivering reagents genome engineering cells. Here, we use geminivirus-based replicons transient expression sequence-specific (zinc-finger nucleases, transcription activator-like effector and the clustered, regularly interspaced, short palindromic repeat/Cas system) delivery DNA repair templates. In tobacco...
Abstract The ability to precisely engineer plant genomes offers much potential for advancing basic and applied biology. Here, we describe methods the targeted modification of using transcription activator-like effector nucleases (TALENs). Methods were optimized tobacco (Nicotiana tabacum) protoplasts TALENs targeting acetolactate synthase (ALS) gene. Optimal TALEN scaffolds identified a protoplast-based single-strand annealing assay in which cleavage creates functional yellow fluorescent...
The zebrafish is a powerful experimental system for uncovering gene function in vertebrate organisms. Nevertheless, studies the have been limited by approaches available eliminating function. Here we present simple and efficient methods inducing, detecting, recovering mutations at virtually any locus zebrafish. Briefly, double-strand DNA breaks are induced of interest synthetic nucleases, called TALENs. Subsequent host repair lesions leads to generation insertion deletion targeted locus. To...
As Caixia Gao and Dan Voytas explain, new techniques make it possible to precisely edit plant genomic DNA, providing opportunities create crop varieties that will help meet the challenges facing agriculture, including an expanding world population environmental change.