25,203 results match your criteria crispr

Genome wide CRISPR screening reveals a role for sialylation in the tumorigenesis and chemoresistance of acute myeloid leukemia cells.

Cancer Lett 2021 Apr 16. Epub 2021 Apr 16.

Department of Convergence Medicine, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea. Electronic address:

Aberrant activation of cytokine and growth factor signal transduction pathways confers enhanced survival and proliferation properties to acute myeloid leukemia (AML) cells. However, the mechanisms underlying the deregulation of signaling pathways in leukemia cells are unclear. To identify genes capable of independently supporting cytokine-independent growth, we employed a genome-wide CRISPR/Cas9-mediated loss-of-function screen in GM-CSF-dependent human AML TF-1 cells. Read More

View Article and Full-Text PDF

High-risk endometrial cancer proteomic profiling reveals that FBXW7 mutation alters L1CAM and TGM2 protein levels.

Cancer 2021 Apr 19. Epub 2021 Apr 19.

Cancer Genetics and Comparative Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland.

Background: FBXW7 is frequently somatically mutated in grade 3 endometrioid endometrial cancers (G3EECs) and serous endometrial cancers (SECs), which are high-risk cancers associated with poor outcomes and in need of novel treatment options. The aim of this study was to determine the proteomic effects of 3 FBXW7 mutations in high-risk endometrial cancers (ECs).

Methods: Clustered regularly interspaced short palindromic repeats (CRISPR) editing was used to generate 3 HEC-50B G3EEC derivative cell lines, each of which harbored 1 FBXW7 mutation, and to revert an endogenous FBXW7 mutation in HEC-1-B grade 2 endometrioid endometrial cancer (G2EEC) cells to the wild-type genotype. Read More

View Article and Full-Text PDF

Population scale nucleic acid delivery to Caenorhabditis elegans via electroporation.

G3 (Bethesda) 2021 Apr 19. Epub 2021 Apr 19.

Alkek Center for Metagenomics and Microbiome Research and Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX 77030, USA.

The free-living nematode C.elegans remains one of the most robust and flexible genetic systems for interrogating the complexities of animal biology. Targeted genetic manipulations, such as RNA interference (RNAi), CRISPR/Cas9- or array-based transgenesis, all depend on initial delivery of nucleic acids. Read More

View Article and Full-Text PDF

Optimising glyphosate tolerance in rapeseed (Brassica napus L.) by CRISPR/Cas9-based geminiviral donor DNA replicon system with Csy4-based single-guide RNA processing.

J Exp Bot 2021 Apr 19. Epub 2021 Apr 19.

National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, China.

Rapeseed (Brassica napus L.) is an important oil crop worldwide, and effective weed control can protect its yields and quality. Farmers can benefit from tolerant cultivars when using herbicides, such as glyphosate. Read More

View Article and Full-Text PDF

The SvFUL2 transcription factor is required for inflorescence determinacy and timely flowering in Setaria viridis.

Plant Physiol 2021 Apr 19. Epub 2021 Apr 19.

Donald Danforth Plant Science Center, Saint Louis, MO, 63132, USA.

Inflorescence architecture in cereal crops directly impacts yield potential through regulation of seed number and harvesting ability. Extensive architectural diversity found in inflorescences of grass species is due to spatial and temporal activity and determinacy of meristems, which control the number and arrangement of branches and flowers, and underlie plasticity. Timing of the floral transition is also intimately associated with inflorescence development and architecture, yet little is known about the intersecting pathways and how they are rewired during development. Read More

View Article and Full-Text PDF

Delivery of the Cas9/sgRNA Ribonucleoprotein Complex in Immortalized and Primary Cells via Virus-like Particles ("Nanoblades").

J Vis Exp 2021 Mar 31(169). Epub 2021 Mar 31.

LBMC, Laboratoire de Biologie et Modélisation de la Cellule Univ Lyon, ENS de Lyon, Université Claude Bernard Lyon 1, CNRS, UMR 5239, INSERM, U1210, Lyon, 69007, France;

The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas system has democratized genome-editing in eukaryotic cells and led to the development of numerous innovative applications. However, delivery of the Cas9 protein and single-guide RNA (sgRNA) into target cells can be technically challenge. Classical viral vectors, such as those derived from lentiviruses (LVs) or adeno-associated viruses (AAVs), allow for efficient delivery of transgenes coding for the Cas9 protein and its associated sgRNA in many primary cells and in vivo. Read More

View Article and Full-Text PDF

Genetic determinants facilitating the evolution of resistance to carbapenem antibiotics.

Elife 2021 Apr 19;10. Epub 2021 Apr 19.

Infectious Diseases and Microbiome Program, The Broad Institute of MIT and Harvard, Cambridge, United States.

In this era of rising antibiotic resistance, in contrast to our increasing understanding of mechanisms that cause resistance, our understanding of mechanisms that influence the propensity to evolve resistance remains limited. Here, we identified genetic factors that facilitate the evolution of resistance to carbapenems, the antibiotic of 'last resort,' in , the major carbapenem resistant species. In clinical isolates, we found that high-level transposon insertional mutagenesis plays an important role in contributing to high-level resistance frequencies in several major and emerging carbapenem-resistant lineages. Read More

View Article and Full-Text PDF

Pridopidine reduces mutant huntingtin-induced endoplasmic reticulum stress by modulation of the Sigma-1 receptor.

J Neurochem 2021 Apr 19. Epub 2021 Apr 19.

The Shmunis School of Biomedicine and Cancer Research, Cell Biology Division, George Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, 69978, Israel.

The endoplasmic reticulum (ER)-localized Sigma-1 receptor (S1R) is neuroprotective in models of neurodegenerative diseases, among them Huntington disease (HD). Recent clinical trials in HD patients and preclinical studies in cellular and mouse HD models suggest a therapeutic potential for the high-affinity S1R agonist pridopidine. However, the molecular mechanisms of the cytoprotective effect are unclear. Read More

View Article and Full-Text PDF

Optimized CRISPR/Cas9-mediated single nucleotide mutation in adherent cancer cell lines.

STAR Protoc 2021 Jun 31;2(2):100419. Epub 2021 Mar 31.

Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University Shanghai Cancer Center, Shanghai Medical College of Fudan University, Shanghai 200032, China.

CRISPR/Cas9 is an efficient, accurate, and optimizable genome-editing tool. Here, we present a modified CRISPR/Cas9 genome-editing protocol for single nucleotide mutation in adherent cell lines. The protocol was adapted to focus on ease of use and efficiency. Read More

View Article and Full-Text PDF

Air Pollution and the Epigenome: A Model Relationship for the Exploration of Toxicoepigenetics.

Curr Opin Toxicol 2017 Oct;6:18-25

National Health and Environmental Effects Research Laboratory, US Environmental Protection Agency, Research Triangle Park, NC 27711.

The field of toxicoepigenetics is rapidly emerging to provide new insights into the relationship between environmental factors, the epigenome, and public health. Toxicoepigenetic data have the potential to revolutionize our understanding of environmental exposure effects and susceptibility. Studies in recent years have demonstrated that exposure to air pollution alters epigenetic modification states; however, continued advancement of the field is limited by the intrinsic complexity of the epigenome and inherent limitations of different types of studies (epidemiological, clinical, and ) that are used in toxicoepigenetics. Read More

View Article and Full-Text PDF
October 2017

Screening neutral sites for metabolic engineering of methylotrophic yeast .

Synth Syst Biotechnol 2021 Jun 31;6(2):63-68. Epub 2021 Mar 31.

Division of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian, 116023, PR China.

Methylotrophic yeast is capable to utilize multiple carbon feedstocks especially methanol as sole carbon source and energy, making it an ideal host for bio-manufacturing. However, the lack of gene integration sites limits its systems metabolic engineering, in particular construction of genome-integrated pathway. We here screened the genomic neutral sites for gene integration without affecting cellular fitness, by genomic integration of an enhanced green fluorescent protein () gene via CRISPR-Cas9 technique. Read More

View Article and Full-Text PDF

Generation of macrophages with altered viral sensitivity from genome-edited rhesus macaque iPSCs to model human disease.

Mol Ther Methods Clin Dev 2021 Jun 17;21:262-273. Epub 2021 Mar 17.

Shin Kaneko Laboratory, Department of Cell Growth and Development, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.

Because of their close biological similarity to humans, non-human primate (NHP) models are very useful for the development of induced pluripotent stem cell (iPSC)-based cell and regenerative organ transplantation therapies. However, knowledge on the establishment, differentiation, and genetic modification of NHP-iPSCs, especially rhesus macaque iPSCs, is limited. We succeeded in establishing iPSCs from the peripheral blood of rhesus macaques (Rh-iPSCs) by combining the Yamanaka reprograming factors and two inhibitors (GSK-3 inhibitor [CHIR 99021] and MEK1/2 inhibitor [PD0325901]) and differentiated the cells into functional macrophages through hematopoietic progenitor cells. Read More

View Article and Full-Text PDF

PRRSV Promotes MARC-145 Cells Entry Into S Phase of the Cell Cycle to Facilitate Viral Replication Degradation of p21 by nsp11.

Front Vet Sci 2021 24;8:642095. Epub 2021 Mar 24.

Key Laboratory of Animal Epidemiology of the Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, China Agricultural University, Beijing, China.

Porcine reproductive and respiratory syndrome virus (PRRSV) remains one of the most economically significant pathogens that seriously affect the global swine industry. Despite sustained efforts, the factors that affect PRRSV replication in host cells are far from being fully elucidated and thus warrants further investigation. In this study, we first demonstrated that PRRSV infection can cause downregulation of endogenous p21 protein in MARC-145 cells in a virus dose-dependent manner. Read More

View Article and Full-Text PDF

LUZP1 Controls Cell Division, Migration and Invasion Through Regulation of the Actin Cytoskeleton.

Front Cell Dev Biol 2021 1;9:624089. Epub 2021 Apr 1.

Center for Cooperative Research in Biosciences (CIC BioGUNE), Basque Research and Technology Alliance, Bizkaia Technology Park, Derio, Spain.

LUZP1 is a centrosomal and actin cytoskeleton-localizing protein that regulates both ciliogenesis and actin filament bundling. As the cytoskeleton and cilia are implicated in metastasis and tumor suppression, we examined roles for LUZP1 in the context of cancer. Here we show that exhibits frequent genomic aberrations in cancer, with a predominance of gene deletions. Read More

View Article and Full-Text PDF

Proteome Analysis of USP7 Substrates Revealed Its Role in Melanoma Through PI3K/Akt/FOXO and AMPK Pathways.

Front Oncol 2021 31;11:650165. Epub 2021 Mar 31.

Department of Anesthesiology, The Affiliated Hospital of Southwest Medical University, Luzhou, China.

The ubiquitin-specific protease 7 (USP7), as a deubiquitinating enzyme, plays an important role in tumor progression by various mechanisms and serves as a potential therapeutic target. However, the functional role of USP7 in melanoma remains elusive. Here, we found that USP7 is overexpressed in human melanoma by tissue microarray. Read More

View Article and Full-Text PDF

Efficient correction of Duchenne muscular dystrophy mutations by SpCas9 and dual gRNAs.

Mol Ther Nucleic Acids 2021 Jun 13;24:403-415. Epub 2021 Mar 13.

Lars Bolund Institute of Regenerative Medicine, Qingdao-Europe Advanced Institute for Life Sciences, BGI-Qingdao, BGI-Shenzhen, Qingdao 266555, China.

CRISPR gene therapy is one promising approach for treatment of Duchenne muscular dystrophy (DMD), which is caused by a large spectrum of mutations in the dystrophin gene. To broaden CRISPR gene editing strategies for DMD treatment, we report the efficient restoration of dystrophin expression in induced myotubes by SpCas9 and dual guide RNAs (gRNAs). We first sequenced 32 deletion junctions generated by this editing method and revealed that non-homologous blunt-end joining represents the major indel type. Read More

View Article and Full-Text PDF

Cloning of human ene in . required the removal of an intragenic Pribnow-Schaller Box before it's Insertion into genomic safe harbor AAVS1 site using CRISPR-Cas9.

F1000Res 2020 23;9:1498. Epub 2020 Dec 23.

Department of Hepatology, Institute of Liver and Biliary Sciences, New Delhi, Delhi, 110070, India.

Genomic safe harbors are sites in the genome which are safe for gene insertion such that the inserted gene will function properly, and the disruption of the genomic location doesn't cause any foreseeable risk to the host. The AAVS1 site is the genetic location which is disrupted upon integration of adeno associated virus (AAV) and is considered a 'safe-harbor' in human genome because about one-third of humans are infected with AAV and so far there is no apodictic evidence that AAV is pathogenic or disruption of AAVS1 causes any disease in man.  Therefore, we chose to target the AAVS1 site for the insertion of , a bile acid transporter which is defective in progressive familial intra hepatic cholestasis type-2 (PFIC-2), a lethal disease of children where cytotoxic bile salts accumulate inside hepatocytes killing them and eventually the patient. Read More

View Article and Full-Text PDF
December 2020

CRISPR/Cas9-Mediated α-ENaC Knockout in a Murine Pancreatic β-Cell Line.

Front Genet 2021 1;12:664799. Epub 2021 Apr 1.

Department of Pathophysiology, Nanjing Medical University, Nanjing, China.

Many ion channels participate in controlling insulin synthesis and secretion of pancreatic β-cells. Epithelial sodium channel (ENaC) expressed in human pancreatic tissue, but the biological role of ENaC in pancreatic β-cells is still unclear. Here, we applied the CRISPR/Cas9 gene editing technique to knockout α-ENaC gene in a murine pancreatic β-cell line (MIN6 cell). Read More

View Article and Full-Text PDF

Pruning and Tending Immune Memories: Spacer Dynamics in the CRISPR Array.

Sandra C Garrett

Front Microbiol 2021 1;12:664299. Epub 2021 Apr 1.

Department of Genetics and Genome Sciences, Institute for Systems Genomics, UConn Health, Farmington, CT, United States.

CRISPR-Cas (Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated genes) is a type of prokaryotic immune system that is unique in its ability to provide sequence-specific adaptive protection, which can be updated in response to new threats. CRISPR-Cas does this by storing fragments of DNA from invading genetic elements in an array interspersed with short repeats. The CRISPR array can be continuously updated through integration of new DNA fragments (termed spacers) at one end, but over time existing spacers become obsolete. Read More

View Article and Full-Text PDF

Gene Silencing Through CRISPR Interference in Bacteria: Current Advances and Future Prospects.

Front Microbiol 2021 31;12:635227. Epub 2021 Mar 31.

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.

Functional genetic screening is an important method that has been widely used to explore the biological processes and functional annotation of genetic elements. CRISPR/Cas (Clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein) is the newest tool in the geneticist's toolbox, allowing researchers to edit a genome with unprecedented ease, accuracy, and high-throughput. Most recently, CRISPR interference (CRISPRi) has been developed as an emerging technology that exploits the catalytically inactive Cas9 (dCas9) and single-guide RNA (sgRNA) to repress sequence-specific genes. Read More

View Article and Full-Text PDF

Generation of VEGF knock-in Cashmere goat via the CRISPR/Cas9 system.

Int J Biol Sci 2021 2;17(4):1026-1040. Epub 2021 Mar 2.

State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, Inner Mongolia University, Hohhot, 010000, China.

Cashmere is a rare and specialised animal fibre, which grows on the outer skin of goats. Owing its low yield and soft, light, and warm properties, it has a high economic value. Here, we attempted to improve existing cashmere goat breeds by simultaneously increasing their fibre length and cashmere yield. Read More

View Article and Full-Text PDF

An all-in-one telomerase assay based on CRISPR-Cas12a trans-cleavage while telomere synthesis.

Anal Chim Acta 2021 May 23;1159:338404. Epub 2021 Mar 23.

The Center for Clinical Molecular Medical Detection, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China. Electronic address:

As one of the crucial factors associated with human life span and cancer progression, telomerase is regarded as an emerging biomarker for cancer diagnosis. Therefore, a facile, rapid and sensitive approach for telomerase activity detection with point-of-care (POC) diagnosis potential is in great demands. Herein, an all-in-one telomerase activity detection assay was established based on the telomere synthesis activated CRISPR-Cas12a system. Read More

View Article and Full-Text PDF

Virus-induced plant volatiles mediate the olfactory behavior of its insect vectors.

Plant Cell Environ 2021 Apr 18. Epub 2021 Apr 18.

State Key Laboratory of Rice Biology & Ministry of Agriculture and Rural Affairs Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou, China.

Plant viruses can manipulate their hosts to release odors that are attractive or repellent to their insect vectors. However, the volatile organic compounds (VOCs), either individually or as mixtures, which play a key role in the olfactory behavior of insect vectors remains largely unknown. Our study focused on green rice leafhoppers (GRLHs) vectoring rice dwarf virus (RDV) revealed that RDV infection significantly induced the emission of (E)-β-caryophyllene and 2-heptanol by rice plants which influenced the olfactory behavior of both non-viruliferous and viruliferous GRLHs. Read More

View Article and Full-Text PDF

A rhomboid-like protease gene from an interspecies translocation confers resistance to cyst nematodes.

New Phytol 2021 Apr 18. Epub 2021 Apr 18.

Plant Breeding Institute, Christian-Albrechts-University of Kiel, Olshausenstrasse 40, D-24098, Kiel, Germany.

Plant-parasitic nematodes are severe pests in crop production worldwide. Chemical control of nematodes has been continuously reduced in the past decades due to environmental and health concerns. Therefore, breeding nematode-resistant crops is an important aim to secure harvests. Read More

View Article and Full-Text PDF

Precision modeling of gall bladder cancer patients in mice based on orthotopic implantation of organoid-derived tumor buds.

Oncogenesis 2021 Apr 17;10(4):33. Epub 2021 Apr 17.

Department of Gastroenterology and Hepatology, Yokohama City University Graduate School of Medicine, Kanagawa, 236-0004, Japan.

Genetically engineered mice (GEM) are the gold standard for cancer modeling. However, strict recapitulation of stepwise carcinogenesis from a single tumor-initiating epithelial cell among genetically intact cells in adults is not feasible with the currently available techniques using GEM. In previous studies, we partially overcame this challenge by physically isolating organs from adult animals, followed by genetic engineering in organoids and subcutaneous inoculation in nude mice. Read More

View Article and Full-Text PDF

Elucidation of metabolic pathways of 25-hydroxyvitamin D3 mediated by Cyp24a1 and Cyp3a using Cyp24a1 knockout rats generated by CRISPR/Cas9 system.

J Biol Chem 2021 Apr 15:100668. Epub 2021 Apr 15.

Department of Pharmaceutical Engineering, Faculty of Engineering, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama 939-0398, Japan. Electronic address:

CYP24A1-deficient (Cyp24a1 KO) rats were generated using the CRISPER/Cas9 system to investigate CYP24A1-dependent or -independent metabolism of 25(OH)D3, the prohormone of calcitriol. Plasma 25(OH)D3 concentrations in Cyp24a1 KO rats were approximately two-fold higher than in wild-type rats. Wild-type rats showed five metabolites of 25(OH)D3 in plasma following oral administration of 25(OH)D3, and these metabolites were not detected in Cyp24a1 KO rats. Read More

View Article and Full-Text PDF

A distinct structure of Cas1-Cas2 complex provides insights into the mechanism for the longer spacer acquisition in Pyrococcus furiosus.

Int J Biol Macromol 2021 Apr 14. Epub 2021 Apr 14.

State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, and Collaborative Innovation Center of Biotherapy, Chengdu 610041, PR China. Electronic address:

In the adaptation stage of CRISPR-Cas systems, the Cas1-Cas2 integrase captures and integrates new invader-derived spacers into the CRISPR locus, serving as a molecular memory of prior infection. As of yet, the structural information of Cas1-Cas2 complex is available only for two species. Here we present the crystal structure of Cas1-Cas2 complex of Pyrococcus furiosus, which showed a distinct architecture from the known Cas1-Cas2 complexes. Read More

View Article and Full-Text PDF

Drosophila NSD deletion induces developmental anomalies similar to those seen in Sotos syndrome 1 patients.

Genes Genomics 2021 Apr 17. Epub 2021 Apr 17.

Department of Biological Sciences, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 05029, Republic of Korea.

Background: Haploinsufficiency of the human nuclear receptor binding suppressor of variegation 3-9, enhancer of zeste, and trithorax (SET) domain 1 (NSD1) gene causes a developmental disorder called Sotos syndrome 1 (SOTOS1), which is associated with overgrowth and macrocephaly. NSD family proteins encoding histone H3 lysine 36 (H3K36) methyltransferases are conserved in many species, and Drosophila has a single NSD homolog gene, NSD.

Objective: To gain insight into the biological functions of NSD1 deficiency in the developmental anomalies seen in SOTOS1 patients using an NSD-deleted Drosophila mutant. Read More

View Article and Full-Text PDF

Peptidylprolyl Isomerase C (Ppic) regulates invariant Natural Killer T cell (iNKT) differentiation in mice.

Eur J Immunol 2021 Apr 17. Epub 2021 Apr 17.

Lymphocyte Development and Leukemogenesis Laboratory, Instituto Gulbenkian de Ciência, Calouste Gulbenkian Foundation, Oeiras, Portugal.

Peptidyl-prolyl cis-trans isomerase C (Ppic) is expressed in several bone marrow hematopoietic progenitors and in T cell precursors. Since the expression profile of Ppic in the hemato-immune system was suggestive that it could play a role in hematopoiesis and/or T lymphocyte differentiation, we sought to test that hypothesis in vivo. Specifically, we generated a Ppic deficient mouse model by targeting the endogenous locus by CRISPR/Cas9 and tested the requirement of Ppic in hematopoiesis. Read More

View Article and Full-Text PDF

NSDHL promotes triple-negative breast cancer metastasis through the TGFβ signaling pathway and cholesterol biosynthesis.

Breast Cancer Res Treat 2021 Apr 16. Epub 2021 Apr 16.

Key Laboratory of Breast Cancer in Shanghai, Department of Breast Surgery, Fudan University Shanghai Cancer Center, 270 Dong-An Road, Build 7, Room 303, Shanghai, 200032, China.

Purpose: Metastasis is the main cause of breast cancer mortality. Recent studies have proved that lipid metabolic reprogramming plays critical roles in breast cancer carcinogenesis and metastasis. We aim to identify critical lipid metabolism genes in breast cancer metastasis. Read More

View Article and Full-Text PDF