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https://www.readbyqxmd.com/read/29453333/scleraxis-is-a-transcriptional-activator-that-regulates-the-expression-of-tenomodulin-a-marker-of-mature-tenocytes-and-ligamentocytes
#1
Chisa Shukunami, Aki Takimoto, Yuriko Nishizaki, Yuki Yoshimoto, Seima Tanaka, Shigenori Miura, Hitomi Watanabe, Tetsushi Sakuma, Takashi Yamamoto, Gen Kondoh, Yuji Hiraki
Tenomodulin (Tnmd) is a type II transmembrane glycoprotein predominantly expressed in tendons and ligaments. We found that scleraxis (Scx), a member of the Twist-family of basic helix-loop-helix transcription factors, is a transcriptional activator of Tnmd expression in tenocytes. During embryonic development, Scx expression preceded that of Tnmd. Tnmd expression was nearly absent in tendons and ligaments of Scx-deficient mice generated by transcription activator-like effector nucleases-mediated gene disruption...
February 16, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29449507/rewritable-multi-event-analog-recording-in-bacterial-and-mammalian-cells
#2
Weixin Tang, David R Liu
We present two CRISPR-mediated analog multi-event recording apparatus (CAMERA) systems that use base editors and Cas9 nucleases to record cellular events in bacteria and mammalian cells. The devices record signal amplitude or duration as changes in the ratio of mutually exclusive DNA sequences (CAMERA 1), or as single-base modifications (CAMERA 2). We achieved recording of multiple stimuli in bacteria or mammalian cells, including exposure to antibiotics, nutrients, viruses, light, and changes in Wnt signaling...
February 15, 2018: Science
https://www.readbyqxmd.com/read/29448632/mpeg-cs-bmi-1rnai-nanoparticles-synthesis-and-its-targeted-inhibition-effect-on-cd133-laryngeal-stem-cells
#3
Xudong Wei, Jian He, Jingyu Wang, Wei Wang
Previous studies have confirmed that CD133+ cells in laryngeal tumor tissue have the characteristics of cancer stem cells. Bmi-1 gene expression is central to the tumorigenicity of CD133+ cells. In this study, we tried to develop a new siRNA carrier system using chitosan-methoxypolyethylene nanoparticles (CS-mPEG-NPs) that exhibit higher tumor-targeting ability and enhanced gene silencing efficacy in CD133+ tumor stem cells. It is hoped to block the self-renewal and kill the stem cells of laryngeal carcinoma...
March 1, 2018: Journal of Nanoscience and Nanotechnology
https://www.readbyqxmd.com/read/29447671/label-free-and-sensitive-microrna-detection-based-on-a-target-recycling-amplification-integrated-superlong-poly-thymine-hosted-copper-nanoparticle-strategy
#4
Fengzhou Xu, Lan Luo, Hui Shi, Xiaoxiao He, Yanli Lei, Jinlu Tang, Dinggeng He, Zhenzhen Qiao, Kemin Wang
Poly(thymine)-hosted copper nanoparticles (poly T-CuNPs) have emerged as a promising label-free fluorophore for bioanalysis, but its application in RNA-related studies is still rarely explored. Herein, by utilizing duplex-specific nuclease (DSN) as a convertor to integrate target recycling mechanism into terminal deoxynucleotidyl transferase (TdT)-mediated superlong poly T-CuNPs platform, a specific and sensitive method for microRNA detection has been developed. In this strategy, a 3'-phosphorylated DNA probe can hybridize with target RNA and then be cut by DSN to produce 3'-hydroxylated fragments, which can be further tailed by TdT with superlong poly T for fluorescent CuNPs synthesis...
June 20, 2018: Analytica Chimica Acta
https://www.readbyqxmd.com/read/29447390/pold3-is-required-for-genomic-stability-and-telomere-integrity-in-embryonic-stem-cells-and-meiosis
#5
Zhongcheng Zhou, Lingling Wang, Feixiang Ge, Peng Gong, Hua Wang, Feng Wang, Lingyi Chen, Lin Liu
Embryonic stem cells (ESCs) and meiosis are featured by relatively higher frequent homologous recombination associated with DNA double strand breaks (DSB) repair. Here, we show that Pold3 plays important roles in DSB repair, telomere maintenance and genomic stability of both ESCs and spermatocytes in mice. By attempting to generate Pold3 deficient mice using CRISPR/Cas9 or transcription activator-like effector nucleases, we show that complete loss of Pold3 (Pold3-/-) resulted in early embryonic lethality at E6...
February 13, 2018: Nucleic Acids Research
https://www.readbyqxmd.com/read/29446996/recent-advances-in-therapeutic-genome-editing-in-china
#6
Yang Yang, Qingnan Wang, Qian Li, Ke Men, Zhiyao He, Hongxin Deng, Weizhi Ji, Yuquan Wei
Editing of the genome to correct disease-causing mutations is a promising approach for the treatment of human diseases. Recent advances in the development of programmable nuclease-based genome editing tools have substantially improved the ability to make precise changes in the human genome. Genome editing technologies are already being used to correct genetic mutations in affected tissues and cells to treat diseases that are refractory to traditional gene therapies. Chinese scientists have made remarkable breakthroughs in the field of therapeutic genome editing, particularly with the first clinical trial involving the clustered regularly interspaced short palindromic repeats-caspase 9 system that began in China...
February 2018: Human Gene Therapy
https://www.readbyqxmd.com/read/29446741/whats%C3%A2-new-in-gene-therapy-of-hemophilia
#7
E Carlos Rodriguez-Merchan
BACKGROUND: Several methods have been investigated to effectively and safely transmit genes that stimulate cells to release therapeutic factor VIII (FVIII) and factor IX (FIX) into the circulation of people with hemophilia (PWH). OBJECTIVE: To review the role of gene therapy (GT) in PWH. METHODS: A Cochrane Library and PubMed (MEDLINE) search related to the role of GT in hemophilia was analyzed. RESULTS: The most promising vectors for hemophilia GT are adeno-associated virus (AAV) and lentivirus...
February 14, 2018: Current Gene Therapy
https://www.readbyqxmd.com/read/29446146/crispr-cas9-mediated-mstn-disruption-and-heritable-mutagenesis-in-goats-causes-increased-body-mass
#8
X Wang, Y Niu, J Zhou, H Zhu, B Ma, H Yu, H Yan, J Hua, X Huang, L Qu, Y Chen
Genetic engineering in livestock has been greatly enhanced through the use of artificial programmed nucleases such as the recently emerged clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system. We recently reported our successful application of the CRISPR/Cas9 system to engineer the goat genome through micro-injection of Cas9 mRNA and sgRNAs targeting MSTN and FGF5 in goat embryos. The phenotypes induced by edited loss-of-function mutations of MSTN remain to be evaluated extensively...
February 2018: Animal Genetics
https://www.readbyqxmd.com/read/29442622/chitosan-stabilized-gold-folate-poly-lactide-co-glycolide-nanoplexes-facilitate-efficient-gene-delivery-in-hepatic-and-breast-cancer-cells
#9
Jude Akinyelu, Moganavelli Singh
The biodegradable polymer, poly(lactide-co-glycolide) is a popular polymer of choice in many nanotherapeutic studies. Herein, we report on the synthesis and evaluation of four chitosan stabilized poly(lactide-co-glycolide) nanoparticles with and without coating with gold, and the targeting ligand, folic acid, as potential non-viral gene delivery vectors. The poly(lactide-co-glycolide) nanoparticles were synthesized via nanoprecipitation/solvent evaporation method in conjunction with the surface functionalizing folic acid and chitosan...
July 1, 2018: Journal of Nanoscience and Nanotechnology
https://www.readbyqxmd.com/read/29442507/functional-insights-revealed-by-the-kinetic-mechanism-of-crispr-cas9
#10
Austin T Raper, Anthony A Stephenson, Zucai Suo
The discovery of prokaryotic adaptive immunity prompted widespread use of the RNA-guided clustered regularly interspaced short palindromic repeat (CRISPR)-associated (Cas) endonuclease Cas9 for genetic engineering. However, its kinetic mechanism remains undefined and details of DNA cleavage poorly characterized. Here, we establish a kinetic mechanism of Streptococcus pyogenes Cas9 from guide-RNA binding through DNA cleavage and product release. Association of DNA to the binary complex of Cas9 and guide-RNA is rate-limiting during the first catalytic turnover, while DNA cleavage from a pre-formed ternary complex of Cas9, guide-RNA, and DNA is rapid...
February 14, 2018: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/29440709/brca1-function-in-the-intra-s-checkpoint-is-activated-by-acetylation-via-a-pcaf-sirt1-axis
#11
Tyler J Lahusen, Seung-Jin Kim, Kai Miao, Zebin Huang, Xiaoling Xu, Chu-Xia Deng
Breast cancer associated gene 1 (BRCA1) function has been shown to be regulated by phosphorylation but the role of acetylation has not been determined. Therefore, we tested whether BRCA1 can be acetylated by the acetyltransferases P300/CBP-associated factor (pCAF), GCN5, and p300. p300 exhibited the highest level of BRCA1 acetylation; however, there was also a decrease in the total level of BRCA1. Therefore, we focused on pCAF and GCN5 because they both acetylated BRCA1 without affecting BRCA1 expression. Further analysis indicated that the acetylated form of BRCA1 is deacetylated by wild-type (WT) SIRT1, but not deacetylase mutant SIRT1, suggesting that SIRT1 is a specific deacetylase of BRCA1...
February 14, 2018: Oncogene
https://www.readbyqxmd.com/read/29440573/the-colm-family-polymorphic-toxins-breaching-the-bacterial-cell-wall
#12
Maarten G K Ghequire, Susan K Buchanan, René De Mot
Bacteria host an arsenal of antagonism-mediating molecules to combat for ecologic space. Bacteriocins represent a pivotal group of secreted antibacterial peptides and proteins assisting in this fight, mainly eliminating relatives. Colicin M, a model for peptidoglycan-interfering bacteriocins in Gram-negative bacteria, appears to be part of a set of polymorphic toxins equipped with such a catalytic domain (ColM) targeting lipid II. Diversifying recombination has enabled parasitism of different receptors and has also given rise to hybrid bacteriocins in which ColM is associated with another toxin module...
February 13, 2018: MBio
https://www.readbyqxmd.com/read/29440496/crispr-cas9-cleavages-in-budding-yeast-reveal-templated-insertions-and-strand-specific-insertion-deletion-profiles
#13
Brenda R Lemos, Adam C Kaplan, Ji Eun Bae, Alexander E Ferrazzoli, James Kuo, Ranjith P Anand, David P Waterman, James E Haber
Harnessing CRISPR-Cas9 technology provides an unprecedented ability to modify genomic loci via DNA double-strand break (DSB) induction and repair. We analyzed nonhomologous end-joining (NHEJ) repair induced by Cas9 in budding yeast and found that the orientation of binding of Cas9 and its guide RNA (gRNA) profoundly influences the pattern of insertion/deletions (indels) at the site of cleavage. A common indel created by Cas9 is a 1-bp (+1) insertion that appears to result from Cas9 creating a 1-nt 5' overhang that is filled in by a DNA polymerase and ligated...
February 13, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29440475/characterization-of-the-molecular-crosstalk-within-the-essential-grc3-las1-pre-rrna-processing-complex
#14
Monica C Pillon, Mack Sobhanhy, Robin E Stanley
Grc3 is an essential well-conserved eukaryotic polynucleotide kinase (PNK) that cooperates with the endoribonuclease Las1 to process the pre-ribosomal RNA (rRNA). Aside from being dependent upon Las1 for coordinated kinase and nuclease function, little is known about Grc3 substrate specificity and the molecular mechanisms governing kinase activity. Here we characterize the kinase activity of Grc3 and identify key similarities and differences between Grc3 and other polynucleotide kinase family members. In contrast to other PNK family members, Grc3 has distinct substrate preferences for RNA substrates in vitro...
February 9, 2018: RNA
https://www.readbyqxmd.com/read/29440319/tudor-staphylococcal-nuclease-is-a-structure-specific-ribonuclease-that-degrades-rna-at-unstructured-regions-during-microrna-decay
#15
Chia-Lung Li, Wei-Zen Yang, Zhonghao Shi, Hanna S Yuan
Tudor staphylococcal nuclease (TSN) is an evolutionarily conserved ribonuclease in eukaryotes that is composed of five staphylococcal nuclease-like domains (SN1 to SN5) and a Tudor domain. TSN degrades hyper-edited double-stranded RNA, including primary miRNA precursors containing multiple I-U and U-I pairs, and mature miRNA during miRNA decay. However, how TSN binds and degrades its RNA substrates remains unclear. Here, we show that the C. elegans TSN (cTSN) is a monomeric Ca 2+ -dependent ribonuclease, cleaving RNA chains at the 5'-side of the phosphodiester linkage to produce degraded fragments with 5'-hydroxyl and 3'-phosphate ends...
February 13, 2018: RNA
https://www.readbyqxmd.com/read/29440262/a-meiotic-xpf-ercc1-like-complex-recognizes-joint-molecule-recombination-intermediates-to-promote-crossover-formation
#16
Arnaud De Muyt, Alexandra Pyatnitskaya, Jessica Andréani, Lepakshi Ranjha, Claire Ramus, Raphaëlle Laureau, Ambra Fernandez-Vega, Daniel Holoch, Elodie Girard, Jérome Govin, Raphaël Margueron, Yohann Couté, Petr Cejka, Raphaël Guérois, Valérie Borde
Meiotic crossover formation requires the stabilization of early recombination intermediates by a set of proteins and occurs within the environment of the chromosome axis, a structure important for the regulation of meiotic recombination events. The molecular mechanisms underlying and connecting crossover recombination and axis localization are elusive. Here, we identified the ZZS (Zip2-Zip4-Spo16) complex, required for crossover formation, which carries two distinct activities: one provided by Zip4, which acts as hub through physical interactions with components of the chromosome axis and the crossover machinery, and the other carried by Zip2 and Spo16, which preferentially bind branched DNA molecules in vitro...
February 9, 2018: Genes & Development
https://www.readbyqxmd.com/read/29439290/cell-death-nucleic-acids-and-immunity-inflammation-beyond-the-grave
#17
REVIEW
Keith B Elkon
Cells of the innate immune system are rigged with sensors that detect nucleic acids derived from microbes, especially viruses. It has become clear that these same sensors that respond to nucleic acids derived from damaged cells or defective intracellular processing are implicated in triggering diseases such as lupus and arthritis. The ways in which cells die and the concomitant presence of proteins and peptides that allow nucleic acids to re-enter cells profoundly influence innate immune responses. This review briefly discusses different types of programmed necrosis such as pyroptosis, necroptosis and NETosis and explains how nucleic acids can engage intracellular receptors and stimulate inflammation...
February 13, 2018: Arthritis & Rheumatology
https://www.readbyqxmd.com/read/29438445/crispr-cas9-in-leishmania-mexicana-a-case-study-of-lmxbtn1
#18
Aygul Ishemgulova, Jana Hlaváčová, Karolina Majerová, Anzhelika Butenko, Julius Lukeš, Jan Votýpka, Petr Volf, Vyacheslav Yurchenko
Leishmania parasites cause human cutaneous, mucocutaneous and visceral leishmaniasis. Several studies proposed involvement of certain genes in infectivity of these parasites based on differential mRNA expression data. Due to unusual gene expression mechanism, functions of such genes must be further validated experimentally. Here, we investigated a role of one of the putative virulence factors, LmxM.22.0010-encoded BTN1 (a protein involved in Batten disease in humans), in L. mexicana infectivity. Due to the incredible plasticity of the L...
2018: PloS One
https://www.readbyqxmd.com/read/29438257/recent-insights-into-vascular-development-from-studies-in-zebrafish
#19
Ryota L Matsuoka, Didier Y R Stainier
PURPOSE OF REVIEW: Zebrafish has provided a powerful platform to study vascular biology over the past 25 years, owing to their distinct advantages for imaging and genetic manipulation. In this review, we summarize recent progress in vascular biology with particular emphasis on vascular development in zebrafish. RECENT FINDINGS: The advent of transcription activator-like effector nuclease and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 genome-editing technologies has dramatically facilitated reverse genetic approaches in zebrafish, as in other models...
February 12, 2018: Current Opinion in Hematology
https://www.readbyqxmd.com/read/29437530/endosomal-escape-of-asos-internalized-by-stabilin-receptors-is-regulated-by-rab5c-and-eea1-during-endosomal-maturation
#20
Colton M Miller, W Brad Wan, Punit P Seth, Edward N Harris
Second-generation (Gen 2) Antisense oligonucleotides (ASOs) show increased nuclease stability and affinity for their RNA targets, which has translated to improved potency and therapeutic index in the clinic. Gen 2 ASOs are typically modified using the phosphorothioate (PS) backbone modification, which enhances ASO interactions with plasma, cell surface, and intracellular proteins. This facilitates ASO distribution to peripheral tissues and also promotes cellular uptake after injection into animals. Previous work identified that Stabilin receptors specifically internalize PS-ASOs in the sinusoidal endothelial cells of the liver and the spleen...
February 13, 2018: Nucleic Acid Therapeutics
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