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https://www.readbyqxmd.com/read/28454397/sphingosine-kinase-1-a-novel-independent-prognosis-biomarker-in-hepatocellular-carcinoma
#1
Huajie Cai, Xuemeng Xie, Ling Ji, Xiaojiao Ruan, Zhihai Zheng
Sphingosine kinase 1 (Sphk1) is an oncogenic kinase that is responsible for the phosphorylation of sphingosine to sphingosine-1-phosphate (S1P). Mounting evidence suggests that Sphk1 serves a crucial role in the proliferation and development of a variety of human cancer cells. However, the role of Sphk1 in hepatocellular carcinoma (HCC) has not been fully elucidated. Therefore, the expression of Sphk1 was examined in 127 formalin-fixed, paraffin-embedded HCC tissues using immunohistochemistry, and its clinical implications and prognostic significance were analyzed...
April 2017: Oncology Letters
https://www.readbyqxmd.com/read/28448636/successful-vitrification-of-pronuclear-stage-pig-embryos-with-a-novel-cryoprotective-agent-carboxylated-%C3%AE%C2%B5-poly-l-lysine
#2
Maki Kamoshita, Tsubasa Kato, Katsuyoshi Fujiwara, Takafumi Namiki, Kazuaki Matsumura, Suong-Hyu Hyon, Junya Ito, Naomi Kashiwazaki
Vitrification is a powerful tool for the efficient production of offspring derived from cryopreserved oocytes or embryos in mammalian species including domestic animals. Genome editing technologies such as transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated (Cas)9 are now available even for domestic species, suggesting that the vitrification of embryos at the pronuclear stage (PN) will be more important because they could provide genomic host cells to be targeted by TALENs or CRISPR/Cas9...
2017: PloS One
https://www.readbyqxmd.com/read/28448066/structural-basis-of-crispr-spycas9-inhibition-by-an-anti-crispr-protein
#3
De Dong, Minghui Guo, Sihan Wang, Yuwei Zhu, Shuo Wang, Zhi Xiong, Jianzheng Yang, Zengliang Xu, Zhiwei Huang
The CRISPR-Cas9 systems are bacterial encoded adaptive immune systems to defend against phages infection, through RNA-guided endonuclease activity of Cas9 to degrade double-stranded DNA bearing a protospacer adjacent motif (PAM) and complementary sequences to the guide RNA1,2,3,4,5. Recently, two anti-CRISPR proteins AcrIIA2 and AcrIIA4 from Listeria monocytogenes (Lmo) prophages have been identified, both of which potently inhibit Streptococcus pyogenes Cas9 (SpyCas9) and LmoCas9 activity in bacteria and human cells6...
April 27, 2017: Nature
https://www.readbyqxmd.com/read/28448034/using-a-fluorescent-pcr-capillary-gel-electrophoresis-technique-to-genotype-crispr-cas9-mediated-knockout-mutants-in-a-high-throughput-format
#4
Muhammad Khairul Ramlee, Jing Wang, Alice M S Cheung, Shang Li
The development of programmable genome-editing tools has facilitated the use of reverse genetics to understand the roles specific genomic sequences play in the functioning of cells and whole organisms. This cause has been tremendously aided by the recent introduction of the CRISPR/Cas9 system-a versatile tool that allows researchers to manipulate the genome and transcriptome in order to, among other things, knock out, knock down, or knock in genes in a targeted manner. For the purpose of knocking out a gene, CRISPR/Cas9-mediated double-strand breaks recruit the non-homologous end-joining DNA repair pathway to introduce the frameshift-causing insertion or deletion of nucleotides at the break site...
April 8, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28446922/corrigendum-building-a-genetic-manipulation-tool-box-for-orchid-biology-identification-of-constitutive-promoters-and-application-of-crispr-cas9-in-the-orchid-dendrobium-officinale
#5
Ling Kui, Haitao Chen, Weixiong Zhang, Simei He, Zijun Xiong, Yesheng Zhang, Liang Yan, Chaofang Zhong, Fengmei He, Junwen Chen, Peng Zeng, Guanghui Zhang, Shengchao Yang, Yang Dong, Wen Wang, Jing Cai
[This corrects the article on p. 2036 in vol. 7, PMID: 28127299.].
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28446681/minute-virus-of-mice-mvm-inhibits-transcription-of-the-cyclin-b1-gene-during-infection
#6
Matthew S Fuller, Kinjal Majumder, David J Pintel
Replication of minute virus of mice (MVM) induces a sustained cellular DNA damage response (DDR) which the virus then exploits to prepare the nuclear environment for effective parvovirus takeover. An essential aspect of the MVM-induced DDR is the establishment of a potent pre-mitotic block, which we previously found to be independent of activated p21 and ATR/Chk1 signaling. This arrest, unlike others previously reported, depends upon the significant, specific depletion of cyclin B1 and its encoding RNA, which precludes cyclin B1/CDK1 complex function thus preventing mitotic entry...
April 26, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28446605/crispr-cas9-mediated-gene-knockout-screens-and-target-identification-via-whole-genome-sequencing-uncover-host-genes-required-for-picornavirus-infection
#7
Heon Seok Kim, Kyungjin Lee, Sangsu Bae, Jeongbin Park, Chong-Kyo Lee, Meehyein Kim, Eunji Kim, Minju Kim, Seokjoong Kim, Chonsaeng Kim, Jin-Soo Kim
Several groups have used genome-wide libraries of lentiviruses encoding small-guide RNAs (sgRNAs) for genetic screens. In most cases, sgRNA expression cassettes are integrated into cells by using lentiviruses, and target genes are statistically estimated by the readout of sgRNA sequences after targeted sequencing. We present a new virus-free method for human gene-knockout screens using a genome-wide library of CRISPR/Cas9 sgRNAs based on plasmids, and target gene identification via whole-genome sequencing (WGS) confirming authentic mutations rather than statistical estimating through targeted amplicon sequencing...
April 26, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28446593/creating-cell-type-specific-mutants-by-enhancer-mutagenesis
#8
Stephen Crews
Cell signaling plays an essential role in development, and knowledge of the identities of the cells sending the signal is critical. This can be a challenge, since signaling pathways and ligands are commonly used at multiple times and in multiple cell types during development. One solution to this problem is to create cell type-specific mutants using CRISPR/Cas9 to mutate enhancers that control different patterns of expression. In this issue of Genes & Development, Rogers and colleagues (pp. 634-638) provide the first use of this method in Drosophila to solve a long-standing issue in patterning of the embryonic central nervous system...
April 1, 2017: Genes & Development
https://www.readbyqxmd.com/read/28446309/-establishment-of-mouse-stably-knockout-of-mysm1-in-msc-cell-line-c3h10t1-2-and-its-effect-on-the-immune-regulation-in-vitro
#9
Yu Liu, Heng Zhu, Xiao-Xia Jiang, Bin Zhang, Li Liao, Yuan-Lin Liu, Yi Zhang, Hu Chen, Ning Mao
OBJECTIVE: To aimed at the establishment of mouse stably knockout of MYSM1 mesenchymal stem cell(MSC) line C3H10T1/2, and to investigate its immunological capacity of MSC in vitro. METHODS: To establish the stably transfected MSC cell line by using CRISPR-Cas9 technology. Then the Flow cytometry, quantitative PCR and Western blot were employed to detect whether the MYSM1 have been knockout yet. Furthermore, the immune modulatory effect of MYSM1(-/-) MSC was tested by addition of MYSM1(-/-) MSC supernatant into spleen lymphocyte and Foxp3 culture...
April 2017: Zhongguo Shi Yan Xue Ye Xue za Zhi
https://www.readbyqxmd.com/read/28445693/high-content-analysis-screening-for-cell-cycle-regulators-using-arrayed-synthetic-crrna-libraries
#10
Žaklina Strezoska, Matthew R Perkett, Eldon T Chou, Elena Maksimova, Emily M Anderson, Shawn McClelland, Melissa L Kelley, Annaleen Vermeulen, Anja van Brabant Smith
The CRISPR-Cas9 system has been utilized for large-scale, loss-of-function screens mainly using lentiviral pooled formats and cell-survival phenotypic assays. Screening in an arrayed format expands the types of phenotypic readouts that can be used to now include high-content, morphology-based assays, and with the recent availability of synthetic crRNA libraries, new studies are emerging. Here, we use a cell cycle reporter cell line to perform an arrayed, synthetic crRNA:tracrRNA screen targeting 169 genes (> 600 crRNAs) and used high content analysis (HCA) to identify genes that regulate the cell cycle...
April 23, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28445541/iqgap1-is-an-oncogenic-target-in-canine-melanoma
#11
Becky H Lee, Poornima H Neela, Michael S Kent, Ashley M Zehnder
Canine oral mucosal melanoma is an aggressive malignant neoplasm and is characterized by local infiltration and a high metastatic potential. The disease progression is similar to that of human oral melanomas. Whereas human cutaneous melanoma is primarily driven by activating mutations in Braf (60%) or Nras (20%), human mucosal melanoma harbors these mutations much less frequently. This makes therapeutic targeting and research modeling of the oral form potentially different from that of the cutaneous form in humans...
2017: PloS One
https://www.readbyqxmd.com/read/28445127/mutation-of-the-sp1-binding-site-in-the-5-flanking-region-of-sry-causes-sex-reversal-in-rabbits
#12
Yuning Song, Tingjun Liu, Yong Wang, Jichao Deng, Mao Chen, Lin Yuan, Yi Lu, Yuxin Xu, Haobin Yao, Zhanjun Li, Liangxue Lai
Sex-determining region Y is a crucial gene that initiates male sex determination in mammals. Mutations of the Sp1-binding site in the 5' flanking region of SRY are associated with clinical male-to-female sex reversal syndrome, although such occurrences are rare and, until now, have not been reported in animal models. In this study, we mutated Sp1-binding sites in the 5' flanking region of the rabbit SRY gene using the CRISPR/Cas9 system. As expected, the SRY-Sp1 knockout rabbits had female external and internal genitalia and exhibited normal female copulatory behaviors, but they were infertile, and the adults displayed reduced follicles...
April 9, 2017: Oncotarget
https://www.readbyqxmd.com/read/28444797/the-transcription-factor-myb115-contributes-to-the-regulation-of-proanthocyanidin-biosynthesis-and-enhances-fungal-resistance-in-poplar
#13
Lijun Wang, Lingyu Ran, Yisu Hou, Qiaoyan Tian, Chaofeng Li, Rui Liu, Di Fan, Keming Luo
Proanthocyanidins (PAs) are major defense phenolic compounds in the leaves of poplar (Populus spp.) in response to abiotic and biotic stresses. Transcriptional regulation of PA biosynthetic genes by the MYB-basic helix-loop-helix (bHLH)-WD40 complexes in poplar is not still fully understood. Here, an Arabidopsis TT2-like gene MYB115 was isolated from Populus tomentosa and characterized by various molecular, genetic and biochemical approaches. MYB115 restored PA productions in the seed coat of the Arabidopsis tt2 mutant...
April 26, 2017: New Phytologist
https://www.readbyqxmd.com/read/28444193/quantitative-assessment-of-timing-efficiency-specificity-and-genetic-mosaicism-of-crispr-cas9-mediated-gene-editing-of-hemoglobin-beta-gene-in-rhesus-monkey-embryos
#14
Uros Midic, Pei-Hsuan Hung, Kailey A Vincent, Benjamin Goheen, Patrick G Schupp, Diane D Chen, Daniel E Bauer, Catherine A VandeVoort, Keith E Latham
Gene editing technologies offer new options for developing novel biomedical research models and for gene and stem cell based therapies. However, applications in many species demand high efficiencies, specificity, and a thorough understanding of likely editing outcomes. To date, overall efficiencies, rates of off-targeting, and degree of genetic mosaicism have not been well-characterized for most species, limiting our ability to optimize methods. As a model gene for measuring these parameters of CRISPR/Cas9 application in a primate species (rhesus monkey), we selected the β-hemoglobin gene (HBB), which also has high relevance to potential application of gene editing and stem-cell technologies for treating human disease...
April 21, 2017: Human Molecular Genetics
https://www.readbyqxmd.com/read/28442604/hepatitis-c-virus-indirectly-disrupts-dna-damage-induced-p53-responses-by-activating-protein-kinase-r
#15
Jonathan K Mitchell, Bentley R Midkiff, Benjamin Israelow, Matthew J Evans, Robert E Lanford, Christopher M Walker, Stanley M Lemon, David R McGivern
Many DNA tumor viruses promote cellular transformation by inactivating the critically important tumor suppressor protein p53. In contrast, it is not known whether p53 function is disrupted by hepatitis C virus (HCV), a unique, oncogenic RNA virus that is the leading infectious cause of liver cancer in many regions of the world. Here we show that HCV-permissive, liver-derived HepG2 cells engineered to constitutively express microRNA-122 (HepG2/miR-122 cells) have normal p53-mediated responses to DNA damage and that HCV replication in these cells potently suppresses p53 responses to etoposide, an inducer of DNA damage, or nutlin-3, an inhibitor of p53 degradation pathways...
April 25, 2017: MBio
https://www.readbyqxmd.com/read/28440293/exploiting-induced-pluripotent-stem-cell-derived-macrophages-to-unravel-host-factors-influencing-chlamydia-trachomatis-pathogenesis
#16
Amy T Y Yeung, Christine Hale, Amy H Lee, Erin E Gill, Wendy Bushell, David Parry-Smith, David Goulding, Derek Pickard, Theodoros Roumeliotis, Jyoti Choudhary, Nick Thomson, William C Skarnes, Gordon Dougan, Robert E W Hancock
Chlamydia trachomatis remains a leading cause of bacterial sexually transmitted infections and preventable blindness worldwide. There are, however, limited in vitro models to study the role of host genetics in the response of macrophages to this obligate human pathogen. Here, we describe an approach using macrophages derived from human induced pluripotent stem cells (iPSdMs) to study macrophage-Chlamydia interactions in vitro. We show that iPSdMs support the full infectious life cycle of C. trachomatis in a manner that mimics the infection of human blood-derived macrophages...
April 25, 2017: Nature Communications
https://www.readbyqxmd.com/read/28440027/the-crispr-cas9-system-their-delivery-in-vivo-and-ex-vivo-applications-and-clinical-development-by-startups
#17
REVIEW
Minjung Song
The CRISPR/Cas9 gene editing system was originally derived from the prokaryotic adaptive immune system mediated by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated proteins (Cas). The system has been successfully applied to genome editing in eukaryotes and has contributed to remarkable advances in the life sciences, in areas ranging from agriculture to genetic disease therapies. For efficient editing and extending the influence of this system, proper delivery of its components is crucial...
April 25, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28439828/interrogation-of-functional-mirna-target-interactions-by-crispr-cas9-genome-engineering
#18
Yale S Michaels, Qianxin Wu, Tudor A Fulga
Post-transcriptional silencing by microRNAs (miRNAs) is a critical constituent of eukaryotic gene regulation. miRNAs are short (~22nt) noncoding RNAs capable of specifically targeting the miRNA-induced-silencing-complex (miRISC) to transcripts bearing a complementary miRNA response element (MRE). Although recent methodological advances have greatly improved our understanding of miRNA biogenesis and the mechanisms by which miRNAs repress their cognate targets, exploring the physiological relevance of direct miRNA-target interactions in vivo has remained an outstanding challenge...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28439815/the-effects-of-voluntary-physical-exercise-activated-neurotrophic-signaling-in-rat-hippocampus-on-mrna-levels-of-downstream-signaling-molecules
#19
Christina A E Solvsten, Tina F Daugaard, Yonglun Luo, Frank de Paoli, Jane H Christensen, Anders L Nielsen
Physical exercise results in the increased expression of neurotrophic factors and the subsequent induction of signal transduction cascades with a positive impact on neuronal functions. In this study, we used a voluntary physical exercise rat model to determine correlations in hippocampus mRNA expression of the neurotropic factors Bdnf, VegfA, and Igf1; their receptors TrkB, Igf1R, VegfR1, and VegfrR2; and downstream signal transducers Creb, Syn1, and Vgf. In hippocampi of physically exercised rats, the mRNA expression levels of Bdnf transcript 4 (Bdnf-t4), VegfA, and Igf1, as well as VegfR1, TrkB, Creb, Vgf, and Syn1, were increased...
April 24, 2017: Journal of Molecular Neuroscience: MN
https://www.readbyqxmd.com/read/28438336/distinct-cis-acting-regions-control-six6-expression-during-eye-field-and-optic-cup-stages-of-eye-formation
#20
Kelley L Ledford, Reyna I Martinez-De Luna, Matthew A Theisen, Karisa D Rawlins, Andrea S Viczian, Michael E Zuber
The eye field transcription factor, Six6, is essential for both the early (specification and proliferative growth) phase of eye formation, as well as for normal retinal progenitor cell differentiation. While genomic regions driving six6 optic cup expression have been described, the sequences controlling eye field and optic vesicle expression are unknown. Two evolutionary conserved regions 5' and a third 3' to the six6 coding region were identified, and together they faithfully replicate the endogenous X. laevis six6 expression pattern...
April 21, 2017: Developmental Biology
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