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PubMed Journals Articles About "Precise Base Editing Acetolactate Synthase Genes Confers Herbicide" RSS

08:01 EST 21st February 2020 | BioPortfolio

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Showing "Precise Base Editing Acetolactate Synthase Genes Confers Herbicide" PubMed Articles 1–25 of 12,000+

Detection and characterisation of resistance to acetolactate synthase inhibiting herbicides in Anisantha and Bromus species in the United Kingdom.

Anisantha and Bromus spp. are widespread and difficult to control, potentially due to the evolution of herbicide resistance. In this study, UK populations of four brome species have been tested for the early development of resistance to acetolactate synthase inhibiting herbicides commonly used in their control.


Increasing the targeting scope and efficiency of base editing with Proxy-BE strategy.

Base editors (BEs) are widely used in precise gene editing due to their simplicity and versatility. However, their efficiencies are hindered by various obstacles. Considering the chromatin microenvironment as a possible obstacle, here, we demonstrate a further development of the proxy-CRISPR strategy, termed Proxy-BE, to increase gene editing efficiency. Specifically, a nuclease-dead Cas9 (dCas9) was bound to the sequence about 20-30 bp away from the target site, potentially improving access to the DNA and,...

Off-target Editing by CRISPR-guided DNA base editors.

Base editing is a genome editing strategy that induces specific single nucleotide changes within genomic DNA. Two major DNA base editors, Cytosine base editors (CBEs) and Adenine base editors (ABEs), have been developed that consist of a Cas9 protein linked to a deaminase enzyme that catalyzes targeted base conversion directed by a sgRNA. This strategy has been used widely for precise genome editing because, unlike CRISPR-Cas nuclease-based genome editing systems, this strategy does not create double strand...


Base Editing in Crops: Current Advances, Limitations, and Future Implications.

Targeted mutagenesis via genome editing technologies holds great promise in developing improved crop varieties to meet future demands. Point mutations or single nucleotide polymorphisms (SNPs) often determine important agronomic traits of crops. Genome editing-based single-base changes could generate elite trait variants in crop plants which help in accelerating crop improvement. Among the genome editing technologies, base editing has emerged as a novel and efficient genome editing approach which enables di...

Improving Editing Efficiency for the Sequences with NGH PAM Using xCas9-Derived Base Editors.

The development of CRISPR/Cas9-mediated base editors (BEs) provided a versatile tool for precise genome editing. The recently developed xCas9-derived base editors (xBEs) that recognize the NG PAM substantially expand the targeting scope in the genome, while their editing efficiency needs to be improved. Here, we described an improved version of xBEs by fusing the BPNLS and Gam to the N terminus of xBEs (BPNLS-Gam-xBE3 and BPNLS-xABE), and this version of base editor displayed higher targeting efficiency for...

Double-check base editing (DBE) for efficient A to G conversions.

With the development of CRISPR/Cas9 technology, a new generation of editing methods that convert specific bases has enabled precise single-base mutations. To date, conversion of cytosine to thymidine and adenine to guanine has been achieved using the cytidine deaminase APOBEC1 and adenosine deaminase (TadA), respectively. However, the base editing efficiency can be unacceptably low in some cell types or at certain target loci. One reason might be the lack of a selective pressure against the survival of non-...

Safeners improve maize tolerance under herbicide toxicity stress by increasing the activity of enzymes in vivo.

Tribenuron-methyl (TM), as one of the sulfonylurea herbicides (SUs), has been widely and effectively applied for many kinds of plants. SUs inhibit plant growth by restraining the biosynthetic pathway of branched-chain amino acids (BCAA) catalyzed by acetolactate synthase (ALS). Safeners are agrochemicals that protect crops from herbicide injuries. To improve the crop tolerance under TM toxicity stress, this paper evaluated the protective effect of N-tosyloxazolidine-3-carboxamide. It was found that most of ...

One Prime for All Editing.

Many targeted base transversions, insertions, and deletions remain challenging due to the lack of precise and efficient genome editing technologies. Recently, Anzalone et al. reported a versatile approach to achieve all types of genome edits, shedding new light on correcting most genetic variants associated with diseases.

Expanding the base editing scope to GA and relaxed NG PAM sites by improved xCas9 system.

Base editors have been developed to be powerful tools to generate precise point mutations. However, their applications are hindered by the strict canonical NGG PAM requirement of Streptococcus pyogenes Cas9 (SpCas9). Cas effectors recognizing different PAMs or relaxed PAMs have been employed to address this limitation. Recently, xCas9 and Cas9-NG were both used to further broaden the editing scope to NG PAM sites (Hu et al., 2018; Nishimasu et al., 2018). xCas9 showed broader PAM recognition including GAA a...

BIG-TREE: Base-Edited Isogenic hPSC Line Generation Using a Transient Reporter for Editing Enrichment.

Current CRISPR-targeted single-nucleotide modifications and subsequent isogenic cell line generation in human pluripotent stem cells (hPSCs) require the introduction of deleterious double-stranded DNA breaks followed by inefficient homology-directed repair (HDR). Here, we utilize Cas9 deaminase base-editing technologies to co-target genomic loci and an episomal reporter to enable single-nucleotide genomic changes in hPSCs without HDR. Together, this method entitled base-edited isogenic hPSC line generation ...

Base editor correction of COL7A1 in recessive dystrophic epidermolysis bullosa patient-derived fibroblasts and iPSCs.

Genome editing represents a promising strategy for therapeutic correction of COL7A1 mutations that cause recessive dystrophic epidermolysis bullosa (RDEB). DNA cleavage followed by homology-directed repair (HDR) using an exogenous template has previously been used to correct COL7A1 mutations. HDR rates can be modest and the double-strand DNA breaks that initiate HDR commonly result in accompanying undesired insertions and deletions (indels). To overcome these limitations, we applied an A•T→G•C adenine...

Simplified adenine base editors improve adenine base editing efficiency in rice.

Adenine base editors (ABEs) have been exploited to introduce targeted adenine (A) to guanine (G) base conversions in various plant genomes, including rice, wheat and Arabidopsis. However, the ABEs reported thus far are all quite inefficient at many target sites in rice, which hampers their applications in plant genome engineering and crop breeding. Here, we show that unlike in the mammalian system, a simplified base editor ABE-P1S (Adenine Base Editor-Plant version 1 Simplified) containing the ecTadA*7.10-n...

Multiplex Gene Disruption by Targeted Base Editing of Yarrowia lipolytica Genome Using Cytidine Deaminase Combined with the CRISPR/Cas9 System.

The oleaginous yeast Yarrowia lipolytica has a tendency to use the non-homologous end joining repair (NHEJ) over the homology directed recombination as double-strand breaks (DSB) repair system, making it difficult to edit the genome using homologous recombination. A recently developed Target-AID (activation-induced cytidine deaminase) base editor, designed to recruit cytidine deaminase (CDA) to the target DNA locus via the CRISPR/Cas9 system, can directly induce C to T mutation without DSB and donor DNA. In...

Engineered Cas9 variant tools expand targeting scope of genome and base editing in rice.

Recently, some newly engineered SpCas9 variants, such as xCas9 3.7 and Cas9-NG, expand the PAM recognition site to NG and show function efficiency in mammalian cells. However, there is still not enough comparative data for their efficiencies on the genome and base editing in plants. In addition, there is also a lack of enhanced specific Cas9 variant with NG PAM. Here, we developed a serial of genome editing tools using different Cas9 variants containing the xCas9, the Cas9-NG and the enhanced specific Cas9-...

Search-and-replace genome editing without double-strand breaks or donor DNA.

Most genetic variants that contribute to disease are challenging to correct efficiently and without excess byproducts. Here we describe prime editing, a versatile and precise genome editing method that directly writes new genetic information into a specified DNA site using a catalytically impaired Cas9 fused to an engineered reverse transcriptase, programmed with a prime editing guide RNA (pegRNA) that both specifies the target site and encodes the desired edit. We performed more than 175 edits in human cel...

Discriminated sgRNAs-based SurroGate System Greatly Enhances the Screening Efficiency of Plant Base-edited Cells.

The development of CRISPR/Cas9-mediated base editing has made genomic modification more efficient. However, selecting genetically modified cells from millions of treated cells, especially plant cells, is still challenging. In this study, an efficient surrogate reporter system was established in rice to enrich base-edited cells based on a defective hygromycin resistance gene. After step-by-step optimization, the Discriminated sgRNAs-based SurroGate system (DisSUGs) was generated by artificially differentiati...

Plant organellar RNA editing: what 30 years of research has revealed.

The central dogma in biology defines the flow of genetic information from DNA to RNA to protein. Accordingly, RNA molecules generally accurately follow the sequences of the genes from which they are transcribed. This rule is transgressed by RNA editing, which creates RNA products that differ from their DNA templates. Analyses of the RNA landscapes of terrestrial plants have indicated that RNA editing (in the form of C-to-U base transitions) is highly prevalent within organelles (i.e., mitochondria and chlor...

Reference gene selection for quantitative real-time PCR (qRT-PCR) expression analysis in Galium aparine L.

To accurately evaluate expression levels of target genes, stable internal reference genes is required for normalization of quantitative real-time PCR (qRT-PCR) data. However, there have been no systematical investigation on the stability of reference genes used in the bedstraw weed, Galium aparine L. (BGA). In this study, the expression profiles of seven traditionally used reference genes, namely 18S, 28S, ACT, GAPDH, EF1α, RPL7 and TBP in BGA were assessed under both biotic (developmental time and tissue)...

Physiological, biochemical and molecular bases of resistance to tribenuron-methyl and glyphosate in Conyza canadensis from olive groves in southern Spain.

Multiple resistance to acetolactate synthase (ALS, EC 2.2.1.6) and 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS, EC 2.5.1.19) inhibitor herbicides was studied in two populations of Conyza canadensis (RTG and STG) harvested in southern Spain. Dose-response and enzymatic activity studies for the ALS-inhibiting herbicides showed only cross-resistance to sulfonylureas group but not to the other ALS chemical groups in the RTG population. Regarding glyphosate, the dose-response studies showed that the RTG p...

Genetic Engineering and Editing of Plants: An Analysis of New and Persisting Questions.

Genetic engineering is a molecular biology technique that enables a gene or genes to be inserted into a plant's genome. The first genetically engineered plants were grown commercially in 1996, and the most common genetically engineered traits are herbicide and insect resistance. Questions and concerns have been raised about the effects of these traits on the environment and human health, many of which are addressed in a pair of 2008 and 2009 articles. As new science is published and new techniques like gen...

Aclonifen targets the solanesyl diphosphate synthase, representing a novel mode of action for herbicides.

Aclonifen is a unique diphenyl ether herbicide. Despite its structural similarities to known inhibitors of the protoporphyrinogen oxidase (e.g., acifluorfen, bifenox or oxadiazon), which result in leaf necrosis, aclonifen causes a different phenotype that is described as bleaching. This is also reflected by the Herbicide Resistance Action Committee (HRAC) classification that categorizes aclonifen as an inhibitor of pigment biosynthesis with an unknown target.

A role for alternative end-joining factors in homologous recombination and genome editing in Chinese hamster ovary cells.

CRISPR technologies greatly foster genome editing in mammalian cells through site-directed DNA double strand breaks (DSBs). However, precise editing outcomes, as mediated by homologous recombination (HR) repair, are typically infrequent and outnumbered by undesired genome alterations. By using knockdown and overexpression studies in Chinese hamster ovary (CHO) cells as well as characterizing repaired DNA junctions, we found that efficient HR-mediated genome editing depends on alternative end-joining (alt-EJ...

AlleleProfileR: A versatile tool to identify and profile sequence variants in edited genomes.

Gene editing strategies, such as zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat/Cas9 (CRISPR/Cas9), are revolutionizing biology. However, quantitative and sensitive detection of targeted mutations are required to evaluate and quantify the genome editing outcomes. Here we present AlleleProfileR, a new analysis tool, written in a combination of R and C++, with the ability to batch process the sequence analysi...

Reduced C-to-U RNA editing rates might play a regulatory role in stress response of Arabidopsis.

C-to-U RNA editing is prevalent in the mitochondrial and chloroplast genes in plants. The C-to-U editing rates are constantly very high. During genome evolution, those edited cytidines are likely to be replaced with thymidines at the DNA level. C-to-U editing events are suggested to be designed for reversing the unfavorable T-to-C DNA mutations. Despite the existing theory showing the importance of editing mechanisms, few studies have investigated the genome-wide adaptive signals of the C-to-U editome or th...

Targeted editing of transcriptional activator MXR1 on the Pichia pastoris genome using CRISPR/Cas9 technology.

A highly efficient and targeted CRISPR/Cas9 gene editing system was constructed for Pichia pastoris (syn Komagataella phaffii). Plasmids containing single guide RNA and the MXR1 homology arms were used to precisely edit the transcriptional activator Mxr1 on the Pichia pastoris (P. pastoris) genome. At the S215 amino acid position of Mxr1, one, two and three nucleotides were precisely deleted or inserted and S215 was also mutated to S215A via a single base substitution. Sequencing of PCR amplicons in the reg...


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