7,203 results match your criteria clustered regularly

Progress of genome editing technology and developmental biology useful for radiation research.

J Radiat Res 2021 May;62(Supplement_1):i53-i63

Department of Disease Model, Research Institute of Radiation Biology and Medicine, Hiroshima University, Hiroshima 734-8553, Japan.

Following the development of genome editing technology, it has become more feasible to create genetically modified animals such as knockout (KO), knock-in, and point-mutated animals. The genome-edited animals are useful to investigate the roles of various functional genes in many fields of biological science including radiation research. Nevertheless, some researchers may experience difficulty in generating genome-edited animals, probably due to the requirement for equipment and techniques for embryo manipulation and handling. Read More

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An exonuclease protection and CRISPR/Cas12a integrated biosensor for the turn-on detection of transcription factors in cancer cells.

Anal Chim Acta 2021 Jun 4;1165:338478. Epub 2021 Apr 4.

School of Nursing, Nanjing Medical University, Nanjing, 211166, China. Electronic address:

Transcription factors (TFs) are critical proteins that regulate the expression of genes, and the abnormal change of TFs levels is directly related to physical dysfunctions. Herein, we developed a clustered regularly interspaced short palindromic repeats (CRISPR)-based biosensor for the measurement of TFs level with the assistance of exonuclease protection assay. A dsDNA (activator) with the ability to activate Cas12a was engineered to contain TFs binding domain, and the binding between TFs and the activator can protect the dsDNA from being digested by exonuclease III (Exo III). Read More

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Bioproduction process of natural products and biopharmaceuticals: Biotechnological aspects.

Biotechnol Adv 2021 May 8:107768. Epub 2021 May 8.

Pharmacognosy Research Laboratories & Herbal Analysis Services UK, University of Greenwich, Chatham-Maritime, Kent ME4 4TB, United Kingdom.

Decades of research have been put in place for developing sustainable routes of bioproduction of high commercial value natural products (NPs) on the global market. In the last few years alone, we have witnessed significant advances in the biotechnological production of NPs. The development of new methodologies has resulted in a better understanding of the metabolic flux within the organisms, which have driven manipulations to improve production of the target product. Read More

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[Structure-based optimization and design of CRISPR protein xCas9].

Sheng Wu Gong Cheng Xue Bao 2021 Apr;37(4):1385-1395

School of Life Sciences, Fudan University, Shanghai 200438, China.

Streptococcus pyogenes Cas9 (SpCas9) has become a powerful genome editing tool, but has a limited range of recognizable protospacer adjacent motifs (PAMs) and shows off-target effects. To address these issues, we present a rational approach to optimize the xCas9 mutant derived from SpCas9 by directed evolution. Firstly, energy minimization with the Rosetta program was applied to optimize the three-dimensional structure of Cas9 to obtain the lowest energy conformation. Read More

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[CRISPR/Cas9 technology in disease research and therapy: a review].

Sheng Wu Gong Cheng Xue Bao 2021 Apr;37(4):1205-1228

College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China.

Genome editing is a genetic manipulation technique that can modify DNA sequences at the genome level, including insertion, knockout, replacement and point mutation of specific DNA fragments. The ultimate principle of genome editing technology relying on engineered nucleases is to generate double-stranded DNA breaks at specific locations in genome and then repair them through non-homologous end joining or homologous recombination. With the intensive study of these nucleases, genome editing technology develops rapidly. Read More

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CRISPR/Cas-Based Epigenome Editing: Advances, Applications, and Clinical Utility.

Trends Biotechnol 2021 May 7. Epub 2021 May 7.

Department of Bioengineering, Rice University, Houston, TX, USA; Department of BioSciences, Rice University, Houston, TX, USA. Electronic address:

The epigenome dynamically regulates gene expression and guides cellular differentiation throughout the lifespan of eukaryotic organisms. Recent advances in clustered regularly interspaced palindromic repeats (CRISPR)/Cas-based epigenome editing technologies have enabled researchers to site-specifically program epigenetic modifications to endogenous DNA and histones and to manipulate the architecture of native chromatin. As a result, epigenome editing has helped to uncover the causal relationships between epigenetic marks and gene expression. Read More

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In vivo CRISPR screening for novel noncoding RNA functional targets in glioblastoma models.

J Neurosci Res 2021 May 10. Epub 2021 May 10.

Department of Neurological Surgery, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.

CRISPR (clustered regularly interspaced short palindromic repeat)-based genetic screens offer unbiased and powerful tools for systematic and specific evaluation of phenotypes associated with specific target genes. CRISPR screens have been utilized heavily in vitro to identify functional coding and noncoding genes in a large number of cell types, including glioblastoma (GB), though no prior study has described the evaluation of CRISPR screening in GB in vivo. Here, we describe a protocol for targeting and transcriptionally repressing GB-specific long noncoding RNAs (lncRNAs) by CRISPR interference (CRISPRi) system in vivo, with tumor growth in the mouse cerebral cortex. Read More

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SEMMs: Somatically Engineered Mouse Models. A New Tool for Disease Modeling for Basic and Translational Research.

Front Oncol 2021 23;11:667189. Epub 2021 Apr 23.

Department of Translational Oncology, Genentech, Inc., South San Francisco, CA, United States.

Most experimental oncology therapies fail during clinical development despite years of preclinical testing rationalizing their use. This begs the question of whether the current preclinical models used for evaluating oncology therapies adequately capture patient heterogeneity and response to therapy. Most of the preclinical work is based on xenograft models where tumor mis-location and the lack of the immune system represent a major limitation for the translatability of many observations from preclinical models to patients. Read More

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CRISPR/Cas9-Induced and Mutations Stacked With Confer High Oleic Acid Seed Oil in Pennycress ( L.).

Front Plant Sci 2021 22;12:652319. Epub 2021 Apr 22.

School of Biological Sciences, Illinois State University, Normal, IL, United States.

Pennycress ( L.) is being domesticated as an oilseed cash cover crop to be grown in the off-season throughout temperate regions of the world. With its diploid genome and ease of directed mutagenesis using molecular approaches, pennycress seed oil composition can be rapidly tailored for a plethora of food, feed, oleochemical and fuel uses. Read More

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Sensitive analysis of single nucleotide variation by Cas13d orthologs, EsCas13d and RspCas13d.

Biotechnol Bioeng 2021 May 8. Epub 2021 May 8.

School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300350, China.

RNA-guided CRISPR (RNA-targeting clustered regularly interspaced short palindromic repeats) effector Cas13d is the smallest class II subtype VI proteins identified so far. Here, two recently identified Cas13d effectors from Eubacterium siraeum (Es) and Ruminococcus sp. (Rsp) were characterized and applied for sensitive nucleic acid detection. Read More

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Genetic diversity and population structure of 'Candidatus Liberibacter asiaticus' associated with citrus Huanglongbing in India based on the prophage types.

World J Microbiol Biotechnol 2021 May 8;37(6):95. Epub 2021 May 8.

Plant Pathology Lab, ICAR-Central Citrus Research Institute, Amravati Road, Nagpur, 440033, India.

Huanglongbing (HLB), also known as 'citrus greening', is an extremely destructive disease of citrus worldwide. HLB is associated with three species of the fastidious proteobacterium, Candidatus Liberibacter asiaticus (CaLas), Ca. L. Read More

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Advances in Clustered, Regularly Interspaced Short Palindromic Repeats (CRISPR)-Based Diagnostic Assays Assisted by Micro/Nanotechnologies.

ACS Nano 2021 May 7. Epub 2021 May 7.

School of Life Sciences, South China Normal University, Guangzhou 510631, China.

Clustered, regularly interspaced short palindromic repeats (CRISPR)-based diagnoses, derived from gene-editing technology, have been exploited for less than 5 years and are now reaching the stage of precommercial use. CRISPR tools have some notable features, such as recognition at physiological temperature, excellent specificity, and high-efficiency signal amplification capabilities. These characteristics are promising for the development of next-generation diagnostic technologies. Read More

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CRISPR-Cas-mediated chromosome engineering for crop improvement and synthetic biology.

Nat Plants 2021 May 6. Epub 2021 May 6.

Botanical Institute, Karlsruhe Institute of Technology, Karlsruhe, Germany.

Plant breeding relies on the presence of genetic variation, as well as on the ability to break or stabilize genetic linkages between traits. The development of the genome-editing tool clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein (Cas) has allowed breeders to induce genetic variability in a controlled and site-specific manner, and to improve traits with high efficiency. However, the presence of genetic linkages is a major obstacle to the transfer of desirable traits from wild species to their cultivated relatives. Read More

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The effect of CRISPR constructs microinjection on the expression of developmental genes in Rag1 knocked-out mice embryo.

Vet Med Sci 2021 May 6. Epub 2021 May 6.

Department of Gametes and Cloning, Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.

Despite all the advances in the production of transgenic mice, the production efficiency of these animal models is still low. Given that the expression of developmental genes has a critical role in growth and development of embryo, we determined the expression pattern of pluripotency, trophectoderm and imprinting genes in the Rag1 (recombination-activating gene 1) knocked-out blastocysts resulting from microinjection of CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) constructs into the zygote cytoplasm of C57bl6 mice. Following microinjection, the embryos were cultured and the gene expression of developed blastocysts and natural blastocysts (Sham and control groups) were evaluated using real-time PCR. Read More

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The glycosyltransferase UGT76B1 modulates N-hydroxy-pipecolic acid homeostasis and plant immunity.

Plant Cell 2021 May;33(3):735-749

Department of Plant Biochemistry, Albrecht-von-Haller-Institute for Plant Sciences, University of Goettingen, D-37077 Goettingen, Germany.

The tradeoff between growth and defense is a critical aspect of plant immunity. Therefore, the plant immune response needs to be tightly regulated. Salicylic acid (SA) is an important plant hormone regulating defense against biotrophic pathogens. Read More

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[Nucleic Acid Delivery across Biological Barriers].

Kanjiro Miyata

Yakugaku Zasshi 2021 ;141(5):635-639

Department of Materials Engineering, Graduate School of Engineering, The University of Tokyo.

One of the current critical issues in nucleic acid delivery is the efficient mRNA delivery into target cells, directed toward clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) genome editing. To this end, we have developed a variety of cationic polyaspartamide derivatives with varying side chain structures because they can form nanocomplexes, termed polyplexes, with mRNA through electrostatic interactions. Interestingly, the delivery functions were highly affected by the chemical structures of the polyaspartamide side chains. Read More

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January 2021

CRISPR-Cas technology based genome editing for modification of salinity stress tolerance responses in rice (Oryza sativa L.).

Mol Biol Rep 2021 May 5. Epub 2021 May 5.

Department of Bioinformatics and Biosciences, Capital University of Science and Technology, Islamabad, Pakistan.

Clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein (Cas) technology is an effective tool for site-specific genome editing, used to precisely induce mutagenesis in different plant species including rice. Salinity is one of the most stressful environmental constraints affecting agricultural productivity worldwide. As plant adaptation to salinity stress is under polygenic control therefore, 51 rice genes have been identified that play crucial role in response to salinity. Read More

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CRISPR screen in cancer: status quo and future perspectives.

Am J Cancer Res 2021 15;11(4):1031-1050. Epub 2021 Apr 15.

Department of Medical Oncology, Jinling Hospital, The First School of Clinical Medicine, Southern Medical University Nanjing 210002, China.

Clustered regularly interspaced short palindromic repeats (CRISPR) system offers a powerful platform for genome manipulation, including protein-coding genes, noncoding RNAs and regulatory elements. The development of CRISPR screen enables high-throughput interrogation of gene functions in diverse tumor biologies, such as tumor growth, metastasis, synthetic lethal interactions, therapeutic resistance and immunotherapy response, which are mostly performed or in transplant models. Recently, direct CRISPR screens have been developed to identify drivers of tumorigenesis in native microenvironment. Read More

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CRISPR Cas System: An efficient tool for cancer modelling.

J Pak Med Assoc 2021 Feb;71(2(B)):718-724

Department of Biotechnology, Abdul Wali Khan University, Mardan, Pakistan.

The Clustered Regularly Interspaced Short Palindromic Repeats-Cas-9 (CRISPR-Cas9) system has been a revolutionising tool in the field of molecular genetics, which provides a versatile range of editing potentials. Researchers can produce breaks or alter genomes with ease using the system. Cancer is one of the multi-gene diseases whose genes need to be studied in detail. Read More

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February 2021

A novel non-viral delivery method that enables efficient engineering of primary human T cells for ex vivo cell therapy applications.

Cytotherapy 2021 Apr 30. Epub 2021 Apr 30.

Avectas Ltd, Maynooth, Ireland. Electronic address:

Background Aims: Next-generation immune cell therapy products will require complex modifications using engineering technologies that can maintain high levels of cell functionality. Non-viral engineering methods have the potential to address limitations associated with viral vectors. However, while electroporation is the most widely used non-viral modality, concerns about its effects on cell functionality have led to the exploration of alternative approaches. Read More

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CRISPR/dCas9 as a Therapeutic Approach for Neurodevelopmental Disorders: Innovations and Limitations Compared to Traditional Strategies.

Dev Neurosci 2021 May 3:1-9. Epub 2021 May 3.

Stem Cell and Neurogenesis Unit, Division of Neuroscience, Ospedale San Raffaele, Milan, Italy.

Brain development is a complex process that requires a series of precise and coordinated events to take place. When alterations in some of those events occur, neurodevelopmental disorders (NDDs) may appear, with their characteristic symptoms, including cognitive, social motor deficits, and epilepsy. While pharmacologic treatments have been the only therapeutic options for many years, more recently the research is turning to the direct removal of the underlying genetic cause of each specific NDD. Read More

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: a potential biocatalyst of renewable chemicals and lignocellulosic ethanol production.

Crit Rev Biotechnol 2021 May 2:1-22. Epub 2021 May 2.

Center of Exact and Technological Sciences - CCET, State University of West Paraná, Cascavel, Brazil.

is an ascomycetous yeast which has shown promising results in cellulosic ethanol and renewable chemicals production. It can survive on a variety of carbon sources under industrially favorable conditions due to its fast growth rate, thermotolerance, and acid tolerance. is generally regarded as a safe (GRAS) microorganism, is widely recognized as a powerhouse for the production of heterologous proteins and is accepted by the US Food and Drug Administration (USFDA) for its pharmaceutical and food applications. Read More

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Patient-Specific induced Pluripotent Stem Cell-Derived Hepatocyte-Like Cells as a Model to Study Autosomal Recessive Hypercholesterolemia.

Stem Cells Dev 2021 May 2. Epub 2021 May 2.

Associatie KU Leuven, 418666, Department of Development and Regeneration, Stem Cell Biology and Embryology, Interdepartmental Stem Cell Institute, Leuven, Belgium;

Autosomal recessive hypercholesterolemia (ARH) is a rare monogenic disorder caused by pathogenic variants in the low-density lipoprotein receptor (LDLR) adaptor protein-1 (LDLRAP1) gene, encoding for the LDLRAP1 protein, which impairs internalization of hepatic LDLR. There are variable responses of ARH patients to treatment and the pathophysiological mechanism(s) for this variability remains unclear. This is in part caused by absence of reliable cellular models to evaluate the effect of LDLRAP1 mutations on LDLRAP1 protein function and its role in LDLR internalization. Read More

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Highly efficient multiplex genetic engineering of porcine primary fetal fibroblasts.

Surg Open Sci 2021 Apr 18;4:26-31. Epub 2020 Nov 18.

Schulze Diabetes Institute, Department of Surgery, University of Minnesota Medical School, Minneapolis, MN, USA.

Background: Genetically engineered porcine donors are a potential solution for the shortage of human organs for transplantation. Incompatibilities between humans and porcine donors are largely due to carbohydrate xenoantigens on the surface of porcine cells, provoking an immune response which leads to xenograft rejection.

Materials And Methods: Multiplex genetic knockout of GGTA1, β4GalNT2, and CMAH is predicted to increase the rate of xenograft survival, as described previously for GGTA1. Read More

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Ubiquitination of Histone H2B by Proteasome Subunit RPT6 Controls Histone Methylation Chromatin Dynamics During Memory Formation.

Biol Psychiatry 2021 Jan 9. Epub 2021 Jan 9.

Department of Neurobiology, University of Alabama at Birmingham, Birmingham, Alabama. Electronic address:

Background: Posttranslational histone modifications play a critical role in the regulation of gene transcription underlying synaptic plasticity and memory formation. One such epigenetic change is histone ubiquitination, a process that is mediated by the ubiquitin-proteasome system in a manner similar to that by which proteins are normally targeted for degradation. However, histone ubiquitination mechanisms are poorly understood in the brain and in learning. Read More

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January 2021

CRISPR-Cas systems: Challenges and future prospects.

Prog Mol Biol Transl Sci 2021 15;180:141-151. Epub 2021 Feb 15.

Department of Biosciences, School of Science, Indrashil University, Rajpur, Mehsana, Gujarat, India. Electronic address:

The advancement gained over the past couple of decades in clustered regularly interspaced short palindromic repeats and CRISPR associated proteins (CRISPR-Cas) systems have revolutionized the field of synthetic biology, therapeutics, diagnostics and metabolic engineering. The technique has enabled the process of genome editing to be very precise, rapid, cost-effective and highly efficient which were the downfalls for the previously debuted zinc finger nucleases (ZFN) and transcription activator-like effector nucleases (TALEN) technologies. However, despite its great potential, challenges including off-target activity, method of delivery, ethical and regulatory issues still remain unresolved for the CRISPR-Cas systems. Read More

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February 2021

CRISPR interference and its applications.

Prog Mol Biol Transl Sci 2021 12;180:123-140. Epub 2021 Feb 12.

Department of Biochemistry and Synthetic Metabolism, Max-Planck Institute for Terrestrial Microbiology, Marburg, Germany. Electronic address:

Sequence-specific control of gene expression is a powerful tool for identifying and studying gene functions and cellular processes. CRISPR interference (CRISPRi) is an RNA-based method for highly specific silencing of the transcription in prokaryotic or eukaryotic cells. The typical CRISPRi system is a type II CRISPR (clustered regularly interspaced palindromic repeats) machinery of Streptococcus pyogenes. Read More

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February 2021

Sensitive detection of foodborne pathogens based on CRISPR-Cas13a.

J Food Sci 2021 Apr 30. Epub 2021 Apr 30.

School of Biology and Biological Engineering, South China University of Technology, Guangzhou, China.

Salmonella, being one of the most widespread foodborne pathogens, is a compulsory test item required by national food safety standard of China and many other countries. More sensitive and specific Salmonella detection method is still needed since traditional methods are time consuming and highly dependent on enormous manpower and material resources. In this research, a bacteria detection method based on CRISPR-Cas13a system (where CRISPR is Clustered Regularly Interspaced Short Palindromic Repeats) was proposed. Read More

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Whole-genome sequencing reveals rare off-target mutations in CRISPR/Cas9-edited grapevine.

Hortic Res 2021 May 1;8(1):114. Epub 2021 May 1.

State Key Laboratory of Crop Stress Biology in Arid Areas, College of Horticulture, Northwest A&F University, 712100, Yangling, Shaanxi, China.

The CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein 9 (Cas9) system is a powerful tool for targeted genome editing, with applications that include plant biotechnology and functional genomics research. However, the specificity of Cas9 targeting is poorly investigated in many plant species, including fruit trees. To assess the off-target mutation rate in grapevine (Vitis vinifera), we performed whole-genome sequencing (WGS) of seven Cas9-edited grapevine plants in which one of two genes was targeted by CRISPR/Cas9 and three wild-type (WT) plants. Read More

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Electroporation-Mediated Genome Editing of Livestock Zygotes.

Front Genet 2021 13;12:648482. Epub 2021 Apr 13.

Department of Animal Science, University of California, Davis, Davis, CA, United States.

The introduction of genome editing reagents into mammalian zygotes has traditionally been accomplished by cytoplasmic or pronuclear microinjection. This time-consuming procedure requires expensive equipment and a high level of skill. Electroporation of zygotes offers a simplified and more streamlined approach to transfect mammalian zygotes. Read More

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