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CRISPR CAS9 human

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https://www.readbyqxmd.com/read/29666172/genetic-editing-of-colonic-organoids-provides-a-molecularly-distinct-and-orthotopic-preclinical-model-of-serrated-carcinogenesis
#1
Tamsin R M Lannagan, Young K Lee, Tongtong Wang, Jatin Roper, Mark L Bettington, Lochlan Fennell, Laura Vrbanac, Lisa Jonavicius, Roshini Somashekar, Krystyna Gieniec, Miao Yang, Jia Q Ng, Nobumi Suzuki, Mari Ichinose, Josephine A Wright, Hiroki Kobayashi, Tracey L Putoczki, Yoku Hayakawa, Simon J Leedham, Helen E Abud, Ömer H Yilmaz, Julie Marker, Sonja Klebe, Pratyaksha Wirapati, Siddhartha Mukherjee, Sabine Tejpar, Barbara A Leggett, Vicki L J Whitehall, Daniel L Worthley, Susan L Woods
OBJECTIVE: Serrated colorectal cancer (CRC) accounts for approximately 25% of cases and includes tumours that are among the most treatment resistant and with worst outcomes. This CRC subtype is associated with activating mutations in the mitogen-activated kinase pathway gene, BRAF , and epigenetic modifications termed the CpG Island Methylator Phenotype, leading to epigenetic silencing of key tumour suppressor genes. It is still not clear which (epi-)genetic changes are most important in neoplastic progression and we begin to address this knowledge gap herein...
April 17, 2018: Gut
https://www.readbyqxmd.com/read/29664675/knockout-of-glucose-transporter-glut6-has-minimal-effects-on-whole-body-metabolic-physiology-in-mice
#2
Frances L Byrne, Ellen M Olzomer, Robert Brink, Kyle L Hoehn
Glucose transporter 6 (GLUT6) is a member of the facilitative glucose transporter family. GLUT6 is up-regulated in several cancers, but is not widely expressed in normal tissues. Previous studies have shown that GLUT6 knockdown kills endometrial cancer cells that express elevated levels of the protein. However, whether GLUT6 represents a viable anti-cancer drug target is unclear because the role of GLUT6 in normal metabolic physiology is unknown. Herein we generated GLUT6 knockout mice to determine how loss of GLUT6 affected whole body glucose homeostasis and metabolic physiology...
April 17, 2018: American Journal of Physiology. Endocrinology and Metabolism
https://www.readbyqxmd.com/read/29663920/advances-in-the-engineering-of-the-gene-editing-enzymes-and-the-genomes-understanding-and-handling-the-off-target-effects-of-crispr-cas9
#3
Yufang Yin, Qian Wang, Li Xiao, Fengjiao Wang, Zhuo Song, Cuilan Zhou, Xuan Liu, Chungen Xing, Nongyue He, Kai Li, Yan Feng, Jia Zhang
In the past decades, significant progresses have been achieved in genetic engineering of nucleases. Among the genetically engineered nucleases, zinc finger nucleases, transcription activator-like (TAL) effector nucleases, and CRIPSPR/Cas9 system form a new field of gene editing. The gene editing efficiency or targeting effect and the off-target effect are the two major determinant factors in evaluating the usefulness of a new enzyme. Engineering strategies in improving these gene editing enzymes, particularly in minimizing their off-target effects, are the focus of this paper...
March 1, 2018: Journal of Biomedical Nanotechnology
https://www.readbyqxmd.com/read/29662178/defining-essential-genes-for-human-pluripotent-stem-cells-by-crispr-cas9-screening-in-haploid-cells
#4
Atilgan Yilmaz, Mordecai Peretz, Aviram Aharony, Ido Sagi, Nissim Benvenisty
The maintenance of pluripotency requires coordinated expression of a set of essential genes. Using our recently established haploid human pluripotent stem cells (hPSCs), we generated a genome-wide loss-of-function library targeting 18,166 protein-coding genes to define the essential genes in hPSCs. With this we could allude to an intrinsic bias of essentiality across cellular compartments, uncover two opposing roles for tumour suppressor genes and link autosomal-recessive disorders with growth-retardation phenotypes to early embryogenesis...
April 16, 2018: Nature Cell Biology
https://www.readbyqxmd.com/read/29662124/stag2-deficiency-induces-interferon-responses-via-cgas-sting-pathway-and-restricts-virus-infection
#5
Siyuan Ding, Jonathan Diep, Ningguo Feng, Lili Ren, Bin Li, Yaw Shin Ooi, Xin Wang, Kevin F Brulois, Linda L Yasukawa, Xingnan Li, Calvin J Kuo, David A Solomon, Jan E Carette, Harry B Greenberg
Cohesin is a multi-subunit nuclear protein complex that coordinates sister chromatid separation during cell division. Highly frequent somatic mutations in genes encoding core cohesin subunits have been reported in multiple cancer types. Here, using a genome-wide CRISPR-Cas9 screening approach to identify host dependency factors and novel innate immune regulators of rotavirus (RV) infection, we demonstrate that the loss of STAG2, an important component of the cohesin complex, confers resistance to RV replication in cell culture and human intestinal enteroids...
April 16, 2018: Nature Communications
https://www.readbyqxmd.com/read/29661693/phosphorylation-of-beta-3-adrenergic-receptor-at-serine-247-by-erk-map-kinase-drives-lipolysis-in-obese-adipocytes
#6
Shangyu Hong, Wei Song, Peter-James H Zushin, Bingyang Liu, Mark P Jedrychowski, Amir I Mina, Zhaoming Deng, Dimitrije Cabarkapa, Jessica A Hall, Colin J Palmer, Hassan Aliakbarian, John Szpyt, Steven P Gygi, Ali Tavakkoli, Lydia Lynch, Norbert Perrimon, Alexander S Banks
OBJECTIVE: The inappropriate release of free fatty acids from obese adipose tissue stores has detrimental effects on metabolism, but key molecular mechanisms controlling FFA release from adipocytes remain undefined. Although obesity promotes systemic inflammation, we find activation of the inflammation-associated Mitogen Activated Protein kinase ERK occurs specifically in adipose tissues of obese mice, and provide evidence that adipocyte ERK activation may explain exaggerated adipose tissue lipolysis observed in obesity...
March 29, 2018: Molecular Metabolism
https://www.readbyqxmd.com/read/29659822/development-and-application-of-crispr-cas9-technologies-in-genomic-editing
#7
Zhang Cui, Quan Renfu, Wang Jinfu
Genomic editing to correct disease-causing mutations is a promising approach for the treatment of human diseases. As a simple and programmable nuclease-based genomic editing tool, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system has substantially improved the ability to make precise changes in the human genome. Rapid development of CRISPR-based technologies in recent years has expanded its application scope and promoted CRISPR-based therapies in preclinical trails...
April 6, 2018: Human Molecular Genetics
https://www.readbyqxmd.com/read/29657966/engineering-complexity-in-bacterial-regulatory-circuits-for-biotechnological-applications
#8
Cauã Antunes Westmann, María-Eugenia Guazzaroni, Rafael Silva-Rocha
Engineering microbial systems allows the generation of new technologies having significant impact in the biotechnological industry and on human health. In the past few years, several synthetic biology approaches have been implemented in bacteria to allow precise engineering of novel regulatory circuits for several applications. The advent of high-throughput technologies and clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-based DNA editing techniques have been pivotal in this process. Yet, despite the tremendous advances experienced recently, there are still a number of bottlenecks that need to be overcome in order to generate high-performance redesigned living machines, and the use of novel computer-aided approaches would be essential for this task...
March 2018: MSystems
https://www.readbyqxmd.com/read/29628935/neurodevelopmental-genetic-diseases-associated-with-microdeletions-and-microduplications-of-chromosome-17p13-3
#9
REVIEW
Sara M Blazejewski, Sarah A Bennison, Trevor H Smith, Kazuhito Toyo-Oka
Chromosome 17p13.3 is a region of genomic instability that is linked to different rare neurodevelopmental genetic diseases, depending on whether a deletion or duplication of the region has occurred. Chromosome microdeletions within 17p13.3 can result in either isolated lissencephaly sequence (ILS) or Miller-Dieker syndrome (MDS). Both conditions are associated with a smooth cerebral cortex, or lissencephaly, which leads to developmental delay, intellectual disability, and seizures. However, patients with MDS have larger deletions than patients with ILS, resulting in additional symptoms such as poor muscle tone, congenital anomalies, abnormal spasticity, and craniofacial dysmorphisms...
2018: Frontiers in Genetics
https://www.readbyqxmd.com/read/29628199/genome-editing-using-crispr-cas9-targeted-mutagenesis-an-opportunity-for-yield-improvements-of-crop-plants-grown-under-environmental-stresses
#10
Mostafa Abdelrahman, Abdullah M Al-Sadi, Alireza Pour-Aboughadareh, David J Burritt, Lam-Son Phan Tran
Developing more crops able to sustainably produce high yields when grown under biotic/abiotic stresses is an important goal, if crop production and food security are to be guaranteed in the face of ever-increasing human population and unpredictable global climatic conditions. However, conventional crop improvement, through random mutagenesis or genetic recombination, is time-consuming and cannot keep pace with increasing food demands. Targeted genome editing (GE) technologies, especially clustered regularly interspaced short palindromic repeats (CRISPR)/(CRISPR)-associated protein 9 (Cas9), have great potential to aid in the breeding of crops that are able to produce high yields under conditions of biotic/abiotic stress...
March 12, 2018: Plant Physiology and Biochemistry: PPB
https://www.readbyqxmd.com/read/29626750/novel-regulatory-roles-of-wnt1-in-infection-associated-colorectal-cancer
#11
Jianwei Wang, Rong Lu, Xinghui Fu, Zhou Dan, Yong-Guo Zhang, Xinxia Chang, Qisha Liu, Yinglin Xia, Xingyin Liu, Jun Sun
Salmonella infection is a major public health concern, and colonization in humans can be chronic and increases the risk of cancers. Wnt signaling is a key pathway for intestinal renewal and development, inflammation, and tumorigenesis. In the current study, we report a novel role of Wnt1 in infection and colon cancer using cell culture models, a Salmonella-colitis colon cancer model, and human samples. In contrast to the bacteria-induced increases in Wnt2 and Wnt11, Salmonella colonization significantly reduced the level of Wnt1 in intestinal epithelial cells in vivo and in vitro...
April 4, 2018: Neoplasia: An International Journal for Oncology Research
https://www.readbyqxmd.com/read/29625575/targeted-genome-engineering-in-human-induced-pluripotent-stem-cells-from-patients-with-hemophilia-b-using-the-crispr-cas9-system
#12
Cuicui Lyu, Jun Shen, Rui Wang, Haihui Gu, Jianping Zhang, Feng Xue, Xiaofan Liu, Wei Liu, Rongfeng Fu, Liyan Zhang, Huiyuan Li, Xiaobing Zhang, Tao Cheng, Renchi Yang, Lei Zhang
BACKGROUND: Replacement therapy for hemophilia remains a lifelong treatment. Only gene therapy can cure hemophilia at a fundamental level. The clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9 (CRISPR-Cas9) system is a versatile and convenient genome editing tool which can be applied to gene therapy for hemophilia. METHODS: A patient's induced pluripotent stem cells (iPSCs) were generated from their peripheral blood mononuclear cells (PBMNCs) using episomal vectors...
April 6, 2018: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/29624775/highly-efficient-ssodn-mediated-homology-directed-repair-of-dsbs-generated-by-cripsr-cas9-in-human-3pn-zygotes
#13
Lichun Tang, Yanting Zeng, Xuewei Zhou, Hongzi Du, Chuang Li, Jianqiao Liu, Pumin Zhang
Ma et al. published data in Nature Journal using human zygotes that showed that double-strand breaks (DSBs) generated by CRISPR/Cas9 could be repaired by homology-directed repair (HDR) using the wild-type allele as a template, by inter-homologue recombination (Ma et al., 2017). This result received great attention from the scientific community, however, in a recent publication, researchers have raised some concerns about Ma's interpretation of their results (Egli et al., 2017). Our previous work showed that ssODN-mediated HDR could repair 75% of DSBs generated by CRISPR/Cas9 at G6PD mutant alleles (Tang et al...
April 6, 2018: Molecular Reproduction and Development
https://www.readbyqxmd.com/read/29622803/simultaneous-reprogramming-and-gene-editing-of-human-fibroblasts
#14
Sara E Howden, James A Thomson, Melissa H Little
The utility of human induced pluripotent stem cells (iPSCs) is enhanced by an ability to precisely modify a chosen locus with minimal impact on the remaining genome. However, the derivation of gene-edited iPSCs typically involves multiple steps requiring lengthy culture periods and several clonal events. Here, we describe a one-step protocol for reliable generation of clonally derived gene-edited iPSC lines from human fibroblasts in the absence of drug selection or FACS enrichment. Using enhanced episomal-based reprogramming and CRISPR/Cas9 systems, gene-edited and passage-matched unmodified iPSC lines are obtained following a single electroporation of human fibroblasts...
May 2018: Nature Protocols
https://www.readbyqxmd.com/read/29622802/design-and-assessment-of-engineered-crispr-cpf1-and-its-use-for-genome-editing
#15
Bin Li, Chunxi Zeng, Yizhou Dong
Cpf1, a CRISPR endonuclease discovered in Prevotella and Francisella 1 bacteria, offers an alternative platform for CRISPR-based genome editing beyond the commonly used CRISPR-Cas9 system originally discovered in Streptococcus pyogenes. This protocol enables the design of engineered CRISPR-Cpf1 components, both CRISPR RNAs (crRNAs) to guide the endonuclease and Cpf1 mRNAs to express the endonuclease protein, and provides experimental procedures for effective genome editing using this system. We also describe quantification of genome-editing activity and off-target effects of the engineered CRISPR-Cpf1 in human cell lines using both T7 endonuclease I (T7E1) assay and targeted deep sequencing...
May 2018: Nature Protocols
https://www.readbyqxmd.com/read/29621903/impact-of-nrf2-on-tumor-growth-and-drug-sensitivity-in-oncogenic-k-ras-transformed-cells-in-vitro-and-in-vivo
#16
Jiajia Shao, Christophe Glorieux, Jianwei Liao, Ping Chen, Wenhua Lu, Zhenhao Liang, Shijun Wen, Yumin Hu, Peng Huang
K-ras is one of the most common oncogenes in human cancers, and its aberrant activation may lead to malignant transformation associated with oxidative stress and activation of the transcription factor Nrf2 that regulates multiple detoxification enzymes. The purpose of this research was to use gene editing technology to evaluate the role of Nrf2 in affecting tumor growth and drug sensitivity of K-rasG12V -transformed cells. We showed that induction of K-rasG12V caused a significant activation of Nrf2 associated with increased expression of its target genes NAD(P)H:quinone oxidoreductase 1 (NQO1) and heme oxygenase-1 (HO-1)...
April 5, 2018: Free Radical Research
https://www.readbyqxmd.com/read/29619162/crispr-cas9-induced-shank3b-mutant-zebrafish-display-autism-like-behaviors
#17
Chun-Xue Liu, Chun-Yang Li, Chun-Chun Hu, Yi Wang, Jia Lin, Yong-Hui Jiang, Qiang Li, Xiu Xu
Background: Human genetic and genomic studies have supported a strong causal role of SHANK3 deficiency in autism spectrum disorder (ASD). However, the molecular mechanism underlying SHANK3 deficiency resulting in ASD is not fully understood. Recently, the zebrafish has become an attractive organism to model ASD because of its high efficiency of genetic manipulation and robust behavioral phenotypes. The orthologous gene to human SHANK3 is duplicated in the zebrafish genome and has two homologs, shank3a and shank3b ...
2018: Molecular Autism
https://www.readbyqxmd.com/read/29617431/membrane-permeabilizing-amphiphilic-peptide-delivers-recombinant-transcription-factor-and-crispr-cas9-cpf1-ribonucleoproteins-in-hard-to-modify-cells
#18
Thomas Del'Guidice, Jean-Pascal Lepetit-Stoffaes, Louis-Jean Bordeleau, Joannie Roberge, Vanessa Théberge, Coraline Lauvaux, Xavier Barbeau, Jessica Trottier, Vibhuti Dave, Denis-Claude Roy, Bruno Gaillet, Alain Garnier, David Guay
Delivery of recombinant proteins to therapeutic cells is limited by a lack of efficient methods. This hinders the use of transcription factors or Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) ribonucleoproteins to develop cell therapies. Here, we report a soluble peptide designed for the direct delivery of proteins to mammalian cells including human stem cells, hard-to-modify primary natural killer (NK) cells, and cancer cell models. This peptide is composed of a 6x histidine-rich domain fused to the endosomolytic peptide CM18 and the cell penetrating peptide PTD4...
2018: PloS One
https://www.readbyqxmd.com/read/29617297/the-combinational-use-of-crispr-cas9-and-targeted-toxin-technology-enables-efficient-isolation-of-bi-allelic-knockout-non-human-mammalian-clones
#19
Satoshi Watanabe, Takayuki Sakurai, Shingo Nakamura, Kazuchika Miyoshi, Masahiro Sato
Recent advances in genome editing systems such as clustered regularly interspaced short palindromic repeats/CRISPR-associated protein-9 nuclease (CRISPR/Cas9) have facilitated genomic modification in mammalian cells. However, most systems employ transient treatment with selective drugs such as puromycin to obtain the desired genome-edited cells, which often allows some untransfected cells to survive and decreases the efficiency of generating genome-edited cells. Here, we developed a novel targeted toxin-based drug-free selection system for the enrichment of genome-edited cells...
April 4, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29615718/genetic-abrogation-of-immune-checkpoints-in-antigen-specific-cytotoxic-t-lymphocyte-as-a-potential-alternative-to-blockade-immunotherapy
#20
Chi Zhang, Yanchun Peng, Philip Hublitz, Haokang Zhang, Tao Dong
T cell function can be compromised during chronic infections or through continuous exposure to tumor antigens by the action of immune checkpoint receptors, such as programmed cell death protein 1 (PD-1). Systemic administration of blocking antibodies against the PD-1 pathway can restore T cell function, and has been approved for the treatment of several malignancies, although there is a risk of adverse immune-related side-effects. We have developed a method for generating gene knockouts in human antigen (Ag)-specific cytotoxic T-Lymphocyte (CTLs) using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome editing...
April 3, 2018: Scientific Reports
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