A5015593109- RNA and protein synthesis mechanisms
- CRISPR and Genetic Engineering
- Plant Reproductive Biology
- RNA Research and Splicing
- Fungal and yeast genetics research
- Plant Virus Research Studies
- Plant Molecular Biology Research
- RNA modifications and cancer
- Chromosomal and Genetic Variations
- Photosynthetic Processes and Mechanisms
- RNA regulation and disease
- Viral Infectious Diseases and Gene Expression in Insects
- Monoclonal and Polyclonal Antibodies Research
- Advanced biosensing and bioanalysis techniques
- Wheat and Barley Genetics and Pathology
- Plant tissue culture and regeneration
- Transgenic Plants and Applications
- Genetically Modified Organisms Research
- Animal Virus Infections Studies
- Insect Resistance and Genetics
- Virus-based gene therapy research
- Genetic Mapping and Diversity in Plants and Animals
- Plant Genetic and Mutation Studies
- Insect symbiosis and bacterial influences
- Peptidase Inhibition and Analysis
Genus (United States)
2019-2024
Pioneer Hi-Bred
2002-2023
Caribou Biosciences (United States)
2023
Corteva (United States)
2020-2022
Eunice Kennedy Shriver National Institute of Child Health and Human Development
1990-1993
National Institutes of Health
1990-1993
Osaka University
1991
Northwestern University
1986-1989
University of Wisconsin–Milwaukee
1986
Abstract Targeted DNA double-strand breaks have been shown to significantly increase the frequency and precision of genome editing. In past two decades, several break technologies developed. CRISPR–Cas9 has quickly become technology choice for editing due its simplicity, efficiency versatility. Currently, in plants primarily relies on delivering reagents form vectors. Here we report biolistic delivery pre-assembled Cas9–gRNA ribonucleoproteins into maize embryo cells regeneration with both...
Targeted mutagenesis, editing of endogenous maize (Zea mays) genes, and site-specific insertion a trait gene using clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas)-guide RNA technology are reported in maize. DNA vectors expressing codon-optimized Streptococcus pyogenes Cas9 endonuclease single guide RNAs were cointroduced with or without repair templates into immature embryos by biolistic transformation targeting five different genomic regions: upstream the...
Recently discovered bacteria and archaea adaptive immune system consisting of clustered regularly interspaced short palindromic repeats (CRISPR) CRISPR-associated (Cas) endonuclease has been explored in targeted genome editing different species. Streptococcus pyogenes Cas9-guide RNA (gRNA) was successfully applied to generate mutagenesis, gene integration, soybean (Glycine max). Two genomic sites, DD20 DD43 on chromosome 4, were mutagenized with frequencies 59% 76%, respectively. Sequencing...
Disease resistance genes in livestock provide health benefits to animals and opportunities for farmers meet the growing demand affordable, high-quality protein. Previously, researchers used gene editing modify porcine CD163 demonstrated a harmful virus that causes reproductive respiratory syndrome (PRRS). To maximize potential benefits, this disease trait needs be present commercially relevant breeding populations multiplication distribution of pigs. Toward goal, first-of-its-kind, scaled...
Phosphorylation of the alpha subunit eukaryotic initiation factor 2 (eIF-2 alpha) in Saccharomyces cerevisiae by GCN2 protein kinase stimulates translation GCN4 mRNA. The kinases heme-regulated inhibitor (HRI) and double-stranded RNA-dependent eIF-2 (dsRNA-PK) inhibit mammalian cells phosphorylating Ser-51 alpha. We show that HRI dsRNA-PK phosphorylate yeast vitro vivo functionally substitute for to stimulate yeast. In addition, high-level expression either decreases growth rate, a finding...
The mechanism by which the scanning ribosome recognizes first AUG codon nearest 5′ end of eukaryotic messenger RNA has not been established. To investigate this an anticodon change (3′-UCC-5′) was introduced into one four methionine initiator (tRNA i met ) genes Saccharomyces cerevisiae . ability mutant transfer to restore growth properties his4 yeast strains in absence histidine then assayed. Only complementary codon, AGG, at region supported His + growth. also directed initiate AGG placed...
Genetic reversion of HIS4 initiator codon mutations in yeast has identified three unlinked genes, sui1, sui2, and SUI3 (suppressors mutations), which when mutated confer the ability to initiate translation at despite absence an AUG start codon. We have previously demonstrated that gene encodes beta subunit eukaryotic initiation factor 2 (eIF-2) a Zn(II) finger motif alter site selection process yeast. In this report, molecular biochemical characterizations show sui2 suppressor alpha eIF-2....
The GCD2 protein is a translational repressor of GCN4, the transcriptional activator multiple amino acid biosynthetic genes in Saccharomyces cerevisiae. We present evidence that has general function initiation synthesis addition to its gene-specific role control GCN4 expression. Two temperature-sensitive lethal gcd2 mutations result sensitivity inhibitors at permissive temperature, and gcd2-503 mutation leads reduced incorporation labeled leucine into total following shift restrictive...
We have developed a novel hybridization platform that utilizes nuclear male sterility to produce hybrids in maize and other cross-pollinating crops. A key component of this is process termed Seed Production Technology (SPT). This incorporates transgenic SPT maintainer line capable propagating nontransgenic male-sterile lines for use as female parents hybrid production. The homozygous recessive sterile transformed with construct containing (i) complementary wild-type fertility gene restore...
Abstract The current rate of yield gain in crops is insufficient to meet the predicted demands. Capturing boost from heterosis one few technologies that offers rapid gain. Hybrids are widely used for cereals, maize and rice, but it has been a challenge develop viable hybrid system bread wheat due genome complexity, which both large hexaploid. Wheat our most grown crop providing 20% calories humans. Here, we describe identification Ms1 , gene proposed use large-scale, low-cost production...
We have mutated various features of the 5' noncoding region HIS4 mRNA in light established Saccharomyces cerevisiae and mammalian consensus translational initiator regions. Our analysis indicates that insertion mutations introduce G + C-rich sequences leader, particularly those result stable stem-loop structures message, severely affect translation initiation. Mutations alter length leader from 115 to 39 nucleotides had no effect on expression, sequence context changes both 3' AUG start...
A unique genetic selection was devised at the HIS4 locus to address mechanism of translation initiation in Saccharomyces cerevisiae and probe sequence requirements normal translational initiator region that might participate ribosomal recognition AUG start codon. The first codon 5' end message serves as site for translation, -3 +4 nucleotide positions flanking this (AXXAUGG) correspond a eucaryotic consensus region. Despite similarity, direct mutations reduce or abolish does not provide any...
GCN4 is a transcriptional activator of amino acid biosynthetic genes in Saccharomyces cerevisiae whose expression regulated by amino-acid availability at the translational level. GCD1 and GCD2 are negative regulators required for repression translation under nonstarvation conditions that mediated upstream open reading frames (uORFs) leader mRNA. GCD factors thought to be antagonized positive GCN1, GCN2 GCN3 acid-starved cells allow increased protein synthesis. Previous genetic studies...
Paramutation involves homologous sequence communication that leads to meiotically heritable transcriptional silencing. We demonstrate mop2 (mediator of paramutation2), which alters paramutation at multiple loci, encodes a gene similar Arabidopsis NRPD2/E2, the second-largest subunit plant-specific RNA polymerases IV and V. In Arabidopsis, Pol-IV Pol-V play major roles in RNA–mediated silencing single is shared between Pol-V. Maize three genes: all genes potentially encode full length...
Modern maize hybrids often contain biotech and native traits. To-date all traits have been randomly inserted in the genome. Consequently, developing with multiple is expensive, time-consuming complex. Here we report using CRISPR-Cas9 to generate a Complex Trait Locus (CTL) facilitate trait stacking. A CTL consists of preselected sites positioned within small well-characterized chromosomal region where genes are inserted. We generated individual lines, each carrying site-specific insertion...
The GCD2 protein is a translational repressor of GCN4, the transcriptional activator multiple amino acid biosynthetic genes in Saccharomyces cerevisiae. We present evidence that has general function initiation synthesis addition to its gene-specific role control GCN4 expression. Two temperature-sensitive lethal gcd2 mutations result sensitivity inhibitors at permissive temperature, and gcd2-503 mutation leads reduced incorporation labeled leucine into total following shift restrictive...
Transcriptional gene silencing has broad applications for studying function in planta. In maize, a large number of genes have been identified as tassel-preferred their expression pattern, both by traditional genetic methods and recent high-throughput profiling platforms. Approaches using RNA suppression may provide rapid alternative means to identify directly related pollen development maize. The male fertility Ms45 several anther-expressed unknown were used evaluate the efficacy generating...
We have mutated various features of the 5' noncoding region HIS4 mRNA in light established Saccharomyces cerevisiae and mammalian consensus translational initiator regions. Our analysis indicates that insertion mutations introduce G + C-rich sequences leader, particularly those result stable stem-loop structures message, severely affect translation initiation. Mutations alter length leader from 115 to 39 nucleotides had no effect on expression, sequence context changes both 3' AUG start...
Targeted mutagenesis using programmable DNA endonucleases has broad applications for studying gene function in planta and developing approaches to improve crop yields. Recently, a genetic method that eliminates the need emasculate female inbred during hybrid seed production, referred as Seed Production Technology, been described. The foundation of this system relied on classical methods identify genes critical anther pollen development. One these is P450 which expressed tapetum anthers....