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https://www.readbyqxmd.com/read/29049737/introduction-of-the-mdm2-t309g-mutation-in-primary-human-retinal-epithelial-cells-enhances-experimental-proliferative-vitreoretinopathy
#1
Guohong Zhou, Yajiang Duan, Gaoen Ma, Wenyi Wu, Zhengping Hu, Na Chen, Yewlin Chee, Jing Cui, Arif Samad, Joanne A Matsubara, Shizuo Mukai, Patricia A D'Amore, Hetian Lei
Purpose: The murine double minute (MDM)2 is a critical negative regulator of the p53 tumor suppressor, and MDM2 SNP309G is associated with a higher risk of proliferative vitreoretinopathy (PVR); in addition, the MDM2 T309G created using clustered regularly interspaced short palindromic repeats (CRISPR)/associated endonuclease (Cas)9 enhances normal rabbit vitreous-induced expression of MDM2 and survival of primary human retinal pigment epithelial (hRPE) cells in vitro. The goal of this study was to determine whether this MDM2 T309G contributes to the development of experimental PVR...
October 1, 2017: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/29046358/translationally-controlled-tumor-protein-is-required-for-the-fast-growth-of-toxoplasma-gondii-and-maintenance-of-its-intracellular-development
#2
Jun Zheng, Yaping Chen, Zhaoran Li, Shinuo Cao, Zhaoxia Zhang, Honglin Jia
Translationally controlled tumor protein (TCTP) is a highly conserved, multifunctional protein that has been implicated in a range of cell physiologic processes, especially cell growth and development. A TCTP-like gene has been identified in the Toxoplasma genome [Toxoplasma gondii TCTP (TgTCTP)], although its function remains unknown. The sequence analysis of TgTCTP indicated that it is a highly conserved protein in eukaryotes. We found that the expression level of TgTCTP in the virulent RH strain was significantly higher than that in the avirulent PLK strain...
October 18, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/29045453/crispr-cas-mediated-knock-in-via-non-homologous-end-joining-in-the-crustacean-daphnia-magna
#3
Hitoshi Kumagai, Takashi Nakanishi, Tomoaki Matsuura, Yasuhiko Kato, Hajime Watanabe
The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated system (Cas) is widely used for mediating the knock-in of foreign DNA into the genomes of various organisms. Here, we report a process of CRISPR/Cas-mediated knock-in via non-homologous end joining by the direct injection of Cas9/gRNA ribonucleoproteins (RNPs) in the crustacean Daphnia magna, which is a model organism for studies on toxicology, ecology, and evolution. First, we confirmed the cleavage activity of Cas9 RNPs comprising purified Cas9 proteins and gRNAs in D...
2017: PloS One
https://www.readbyqxmd.com/read/29039455/cell-penetrating-peptides-and-their-utility-in-genome-function-modifications-review
#4
Maciej Gagat, Wioletta Zielińska, Alina Grzanka
For almost 30 years, studies have confirmed the effectiveness of cell-penetrating peptides (CPPs) in the facilitation of the intracellular delivery of various cargo molecules, including RNA, DNA, plasmids, proteins or nanoparticles, under in vitro and in vivo conditions. The cellular uptake of CPPs occurs via energy-dependent, as well as -independent mechanisms. In this relatively new direction of research, studies have attempted to introduce genome modification systems into cells by CPPs. Cellular uptake of CPPs carrying either covalently bound or electrostatically conjugated cargo, has several advantages over viral delivery systems, as it does not lead to any significant cytotoxicity or immunogenicity, and simultaneously it is more efficient than other non-viral systems...
October 4, 2017: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/29038911/negative-regulation-of-daptomycin-production-by-depr2-an-arsr-family-transcriptional-factor
#5
Xu-Ming Mao, Shuai Luo, Yong-Quan Li
Daptomycin, a lipopeptide antibiotic potently active against Gram-positive bacterial pathogens, is produced by Streptomyces roseosporus, but the transcriptional regulation on its biosynthesis is not fully understood. Here, we report that DepR2, an ArsR-family transcriptional regulator isolated previously by DNA-affinity purification, interacts directly with dptEp, the major promoter of the daptomycin gene cluster. DepR2 binds to an imperfect palindromic sequence at the very upstream of dptEp. Meanwhile, higher dptEp activities were consistently observed in the ΔdepR2 mutant, correlating with a nearly 2...
October 16, 2017: Journal of Industrial Microbiology & Biotechnology
https://www.readbyqxmd.com/read/29038257/the-novel-transcriptional-regulator-lmbu-promotes-lincomycin-biosynthesis-through-regulating-expression-of-its-target-genes-in-streptomyces-lincolnensis
#6
Bingbing Hou, Yanwei Lin, Haizhen Wu, Meijin Guo, Hrvoje Petkovic, Liyuan Tao, Xiaoyu Zhu, Jiang Ye, Huizhan Zhang
Lincomycin A is a clinically important antimicrobial agent produced by Streptomyces lincolnensis In this study, a new regulator designated LmbU (Genbank accession no. ABX00623.1) was identified and characterized to regulate lincomycin biosynthesis in a S. lincolnensis wild-type strain NRRL 2936. Both inactivation and overexpression of lmbU result in significant influences on lincomycin production. Transcriptional analysis and in vivo neomycin resistance (neo(r) ) reporter assays demonstrated that LmbU activates expression of the lmbA, lmbC, lmbJ and lmbW genes, and represses expression of the lmbK and lmbU genes...
October 16, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/29035497/conformational-dynamics-of-dna-binding-and-cas3-recruitment-by-the-crispr-rna-guide-cascade-complex
#7
Paul B G van Erp, Angela Patterson, Ravi Kant, Luke Berry, Sarah M Golden, Brittney L Forsman, Joshua Carter, Ryan N Jackson, Brian Bothner, Blake Wiedenheft
Bacteria and archaea rely on CRISPR (clustered regularly interspaced short palindromic repeats) RNA-guided adaptive immune systems for sequence specific elimination of foreign nucleic acids. In Escherichia coli, short CRISPR-derived RNAs (crRNAs) assemble with Cas (CRISPR-associated) proteins into a 405-kilodalton multi-subunit surveillance complex called Cascade (CRISPR-associated complex for antiviral defense). Cascade binds foreign DNA complementary to the crRNA guide and recruits Cas3, a trans-acting nuclease-helicase required for target degradation...
October 16, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/29032652/molecular-characterization-of-salmonella-genomic-island-1-in-proteus-mirabilis-isolates-from-chungcheong-provinces-korea
#8
Ji Youn Sung, Semi Kim, GyeCheol Kwon, Sun Hoe Koo
Emergence and dissemination of Salmonella genomic island 1 (SGI1) is strongly associated with occurrence of multidrug resistant (MDR) Enterobacteria in humans and animals. Diverse SGI1s have been reported among Salmonella enterica and Proteus mirabilis in several countries. We aimed to characterize SGI1 in P. mirabilis isolates from humans and chickens at ChungCheong provinces, Korea. A total of 44 P. mirabilis isolates were recovered from humans (n=20) and chickens (n=24). Antimicrobial susceptibility was determined by disk diffusion method...
October 14, 2017: Journal of Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29028415/crispr-cas9-mediated-noncoding-rna-editing-in-human-cancers
#9
Jie Yang, Xiaodan Meng, Jinchang Pan, Nan Jiang, Chengwei Zhou, Zhenhua Wu, Zhaohui Gong
Cancer is characterized by multiple genetic and epigenetic alterations, including a higher prevalence of mutations of oncogenes and/or tumor suppressors. Mounting evidences have shown that noncoding RNAs (ncRNAs) are involved in the epigenetic regulation of cancer genes and their associated pathways. The clustered regularly interspaced short palindromic repeats (CRISPR)-associated nuclease 9 (CRISPR/Cas9) system, a revolutionary genome-editing technology, has shed light on ncRNA-based cancer therapy. Here, we briefly introduce the classifications and mechanisms of CRISPR/Cas9 system...
October 13, 2017: RNA Biology
https://www.readbyqxmd.com/read/29028182/hypothesis-rna-and-dna-viral-sequence-integration-into-the-mammalian-host-genome-supports-long-term-b-cell-and-t-cell-adaptive-immunity
#10
Julia L Hurwitz, Bart G Jones, Emmanuelle Charpentier, David L Woodland
Viral sequence integration into the mammalian genome has long been perceived as a health risk. In some cases, integration translates to chronic viral infection, and in other instances, oncogenic gene mutations occur. However, research also shows that animal cells can benefit from integrated viral sequences (e.g., to support host cell development or to silence foreign invaders). Here we propose that, comparable with the clustered regularly interspaced short palindromic repeats that provide bacteria with adaptive immunity against invasive bacteriophages, animal cells may co-opt integrated viral sequences to support immune memory...
October 13, 2017: Viral Immunology
https://www.readbyqxmd.com/read/29026866/a-prospective-observational-study-of-the-epidemiology-management-and-outcomes-of-skin-and-soft-tissue-infections-due-to-carbapenem-resistant-enterobacteriaceae
#11
Oryan Henig, Eric Cober, Sandra S Richter, Federico Perez, Robert A Salata, Robert C Kalayjian, Richard R Watkins, Steve Marshall, Susan D Rudin, T Nicholas Domitrovic, Andrea M Hujer, Kristine M Hujer, Yohei Doi, Scott Evans, Vance G Fowler, Robert A Bonomo, David van Duin, Keith S Kaye
BACKGROUND: This study was performed to characterize the epidemiology, management, and outcomes of skin and soft tissue infection (SSTI) and colonization due to carbapenem-resistant Enterobacteriaceae (CRE). METHODS: Patients from the Consortium on Resistance Against Carbapenem in Klebsiella and Other Enterobacteriaceae (CRACKLE-1) from December 24, 2011 to October 1, 2014 with wound cultures positive for CRE were included in the study. Predictors of surgical intervention were analyzed...
2017: Open Forum Infectious Diseases
https://www.readbyqxmd.com/read/29026656/phylogenomics-and-comparative-genomics-of-lactobacillus-salivarius-a-mammalian-gut-commensal
#12
Hugh M B Harris, Maxence J B Bourin, Marcus J Claesson, Paul W O'Toole
The genus Lactobacillus is a diverse group with a combined species count of over 200. They are the largest group within the lactic acid bacteria and one of the most important bacterial groups involved in food microbiology and human nutrition because of their fermentative and probiotic properties. Lactobacillus salivarius, a species commonly isolated from the gastrointestinal tract of humans and animals, has been described as having potential probiotic properties and results of previous studies have revealed considerable functional diversity existing on both the chromosomes and plasmids...
August 2017: Microbial Genomics
https://www.readbyqxmd.com/read/29024560/enhanced-stem-cell-differentiation-and-immunopurification-of-genome-engineered-human-retinal-ganglion-cells
#13
Valentin M Sluch, Xitiz Chamling, Melissa M Liu, Cynthia A Berlinicke, Jie Cheng, Katherine L Mitchell, Derek S Welsbie, Donald J Zack
Human pluripotent stem cells have the potential to promote biological studies and accelerate drug discovery efforts by making possible direct experimentation on a variety of human cell types of interest. However, stem cell cultures are generally heterogeneous and efficient differentiation and purification protocols are often lacking. Here, we describe the generation of clustered regularly-interspaced short palindromic repeats(CRISPR)-Cas9 engineered reporter knock-in embryonic stem cell lines in which tdTomato and a unique cell-surface protein, THY1...
October 10, 2017: Stem Cells Translational Medicine
https://www.readbyqxmd.com/read/29021165/preclinical-modeling-highlights-the-therapeutic-potential-of-hematopoietic-stem-cell-gene-editing-for-correction-of-scid-x1
#14
Giulia Schiroli, Samuele Ferrari, Anthony Conway, Aurelien Jacob, Valentina Capo, Luisa Albano, Tiziana Plati, Maria C Castiello, Francesca Sanvito, Andrew R Gennery, Chiara Bovolenta, Rahul Palchaudhuri, David T Scadden, Michael C Holmes, Anna Villa, Giovanni Sitia, Angelo Lombardo, Pietro Genovese, Luigi Naldini
Targeted genome editing in hematopoietic stem/progenitor cells (HSPCs) is an attractive strategy for treating immunohematological diseases. However, the limited efficiency of homology-directed editing in primitive HSPCs constrains the yield of corrected cells and might affect the feasibility and safety of clinical translation. These concerns need to be addressed in stringent preclinical models and overcome by developing more efficient editing methods. We generated a humanized X-linked severe combined immunodeficiency (SCID-X1) mouse model and evaluated the efficacy and safety of hematopoietic reconstitution from limited input of functional HSPCs, establishing thresholds for full correction upon different types of conditioning...
October 11, 2017: Science Translational Medicine
https://www.readbyqxmd.com/read/29019352/hot-news-gene-therapy-with-crispr-cas9-coming-to-age-for-hiv-cure
#15
Vicente Soriano
The huge success of current antiretroviral therapy is mediated by a triple effect: (i) Halting progression to AIDS in infected persons; (ii) reducing the risk of transmission to contacts (treatment as prevention); and (iii) minimizing the risk of HIV acquisition treating uninfected persons at risk (pre-exposure prophylaxis). However, UNAIDS has estimated that only 70% of infected people globally are diagnosed, only 53% are treated, and overall 44% have undetectable viral load, which is the necessary request for ensuring any antiretroviral benefit...
October 11, 2017: AIDS Reviews
https://www.readbyqxmd.com/read/29017452/whole-genome-sequencing-for-the-molecular-characterization-of-carbapenem-resistant-klebsiella-pneumoniae-strains-isolated-at-the-italian-asst-fatebenefratelli-sacco-hospital-2012-2014
#16
Sara Giordana Rimoldi, Bernardina Gentile, Cristina Pagani, Annamaria Di Gregorio, Anna Anselmo, Anna Maria Palozzi, Antonella Fortunato, Valentina Pittiglio, Anna Lisa Ridolfo, Maria Rita Gismondo, Giuliano Rizzardini, Florigio Lista
BACKGROUND: The emergence of carbapenem-resistant Klebsiella pneumoniae strains is threatening antimicrobial treatment. METHODS: Sixty-eight carbapenemase-producing K. pneumoniae strains isolated at Luigi Sacco University Hospital-ASST Fatebenefratelli Sacco (Milan, Italy) between 2012 and 2014 were characterised microbiologically and molecularly. They were tested for drug susceptibility and carbapenemase phenotypes, investigated by means of repetitive extra-genic palindromic polymerase chain reaction (REP-PCR), and fully sequenced by means of next-generation sequencing for the in silico analysis of multi-locus sequence typing (MLST), their resistome, virulome and plasmid content, and their core single nucleotide polymorphism (SNP) genotypes...
October 10, 2017: BMC Infectious Diseases
https://www.readbyqxmd.com/read/29016293/molecular-and-phenotypic-characterization-of-diarrheagenic-escherichia-coli-strains-isolated-from-bacteremic-children
#17
Maribel Riveros, Wilfredo García, Coralith García, David Durand, Erik Mercado, Joaquim Ruiz, Theresa J Ochoa
Escherichia coli is an important cause of Gram-negative bacteremia. The aim of this study was to characterize at the molecular and phenotypic levels E. coli strains belonging to different diarrheagenic pathotypes [diarrheagenic E. coli (DEC)] isolated from bacteremia in children younger than 5 years of age. Seventy bacteremia E. coli strains were collected in a prospective study in 12 hospitals in Lima, Peru. The presence of virulence genes associated with DEC [enterotoxigenic (lt and st), enteropathogenic (eaeA), shiga toxin-producing (stx1and stx2), enteroinvasive (ipaH), enteroaggregative (aggR), and diffusely adherent (daaD)] was determined by multiplex real-time polymerase chain reaction (PCR)...
October 9, 2017: American Journal of Tropical Medicine and Hygiene
https://www.readbyqxmd.com/read/28994414/mismodeled-purines-implicit-alternates-and-hidden-hoogsteens
#18
Bradley J Hintze, Jane S Richardson, David C Richardson
Hoogsteen base pairs are seen in DNA crystal structures, but only rarely. This study tests whether Hoogsteens or other syn purines are either under-modeled or over-modeled, which are known problems for rare conformations. Candidate purines needing a syn/anti 180° flip were identified by diagnostic patterns of difference electron-density peaks. Manual inspection narrowed 105 flip candidates to 20 convincing cases, all at ≤2.7 Å resolution. Rebuilding and refinement confirmed that 14 of these were authentic purine flips...
October 1, 2017: Acta Crystallographica. Section D, Structural Biology
https://www.readbyqxmd.com/read/28992411/genome-editing-insights-from-chemical-biology-to-support-safe-and-transformative-therapeutic-applications
#19
Renee D Wegrzyn, Andrew H Lee, Amy L Jenkins, Colby D Stoddard, Anne E Cheever
Programmable nuclease-based genome editing technologies, including the clustered, regularly interspaced, short palindromic repeats (CRISPR)/Cas9 system, are becoming an essential component of many applications ranging from agriculture to medicine. However, fundamental limitations including the potential for off-target effects, limited control of editing activity and subsequent DNA repair outcomes, and insufficient target conversion and delivery performance prevent the widespread, safe, and practical use of genome editors, especially for human disease interventions...
October 9, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28988229/studying-kidney-disease-using-tissue-and-genome-engineering-in-human-pluripotent-stem-cells
#20
Elena Garreta, Federico González, Núria Montserrat
Kidney morphogenesis and patterning have been extensively studied in animal models such as the mouse and zebrafish. These seminal studies have been key to define the molecular mechanisms underlying this complex multistep process. Based on this knowledge, the last 3 years have witnessed the development of a cohort of protocols allowing efficient differentiation of human pluripotent stem cells (hPSCs) towards defined kidney progenitor populations using two-dimensional (2D) culture systems or through generating organoids...
October 7, 2017: Nephron
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