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Antisense oligos

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https://www.readbyqxmd.com/read/27860312/cytochrome-p450-26a1-modulates-natural-killer-cells-in-mouse-early-pregnancy
#1
Chao-Yang Meng, Zhong-Yin Li, Wen-Ning Fang, Zhi-Hui Song, Dan-Dan Yang, Dan-Dan Li, Ying Yang, Jing-Pian Peng
Cytochrome P450 26A1 (CYP26A1) has a spatiotemporal expression pattern in the uterus, with a significant increase in mRNA and protein levels during peri-implantation. Inhibiting the function or expression of CYP26A1 can cause pregnancy failure, suggesting an important regulatory role of CYP26A1 in the maintenance of pregnancy. However, little is known about the exact mechanism involved. In this study, using a pCR3.1-cyp26a1 plasmid immunization mouse model and a Cyp26a1-MO (Cyp26a1-specific antisense oligos) knockdown mouse model, we report that the number of Dolichos biflorus agglutinin (DBA) lectin-positive uterine natural killer (uNK) cells was reduced in pCR3...
November 17, 2016: Journal of Cellular and Molecular Medicine
https://www.readbyqxmd.com/read/27830347/mito-porter-for-mitochondrial-delivery-and-mitochondrial-functional-analysis
#2
Yuma Yamada, Hideyoshi Harashima
Mitochondria are attractive organelles that have the potential to contribute greatly to medical therapy, the maintenance of beauty and health, and the development of the life sciences. Therefore, it would be expected that the further development of mitochondrial drug delivery systems (DDSs) would exert a significant impact on the medical and life sciences. To achieve such an innovative objective, it will be necessary to deliver various cargoes to mitochondria in living cells. However, only a limited number of approaches are available for accomplishing this...
November 10, 2016: Handbook of Experimental Pharmacology
https://www.readbyqxmd.com/read/27797146/innovative-technologies-in-nanomedicines-from-passive-targeting-to-active-targeting-from-controlled-pharmacokinetics-to-controlled-intracellular-pharmacokinetics
#3
Yusuke Sato, Yu Sakurai, Kazuaki Kajimoto, Takashi Nakamura, Yuma Yamada, Hidetaka Akita, Hideyoshi Harashima
Nanomedicines promise to extend drug therapy from small molecular compounds to proteins/nucleic acids/genes. Multifunctional envelope-type nanodevices (MENDs) have been developed for delivering such molecules to the site of action. The YSK-MEND contains new types of pH-responsive cationic lipids to efficiently deliver siRNA to hepatocytes via receptor-mediated endocytosis and use in treating hepatitis C and B in model mice. The RGD ligand is introduced to target tumor endothelial cells (TEC) and RGD-MEND is able to send siRNA to TEC to regulate the function of tumor microenvironments...
October 31, 2016: Macromolecular Bioscience
https://www.readbyqxmd.com/read/27792754/characterization-of-zebrafish-models-of-marinesco-sj%C3%A3-gren-syndrome
#4
Genri Kawahara, Yukiko K Hayashi
SIL1 is a nucleotide exchange factor for the endoplasmic reticulum chaperone, BiP. Mutations in the SIL1 gene cause Marinesco-Sjögren syndrome (MSS), an autosomal recessive disease characterized by cerebellar ataxia, mental retardation, congenital cataracts, and myopathy. To create novel zebrafish models of MSS for therapeutic drug screening, we analyzed phenotypes in sil1 knock down fish by two different antisense oligo morpholinos. Both sil1 morphants had abnormal formation of muscle fibers and irregularity of the myosepta...
2016: PloS One
https://www.readbyqxmd.com/read/27746303/a-versatile-tandem-rna-isolation-procedure-to-capture-in-vivo-formed-mrna-protein-complexes
#5
Ana M Matia-González, Valentina Iadevaia, André P Gerber
We describe a tandem RNA isolation procedure (TRIP) that enables purification of in vivo formed messenger ribonucleoprotein (mRNP) complexes. The procedure relies on the purification of polyadenylated mRNAs with oligo(dT) beads from cellular extracts, followed by the capture of specific mRNAs with 3'-biotinylated 2'-O-methylated antisense RNA oligonucleotides, which are recovered with streptavidin beads. TRIP was applied to isolate in vivo crosslinked mRNP complexes from yeast, nematodes and human cells for subsequent analysis of RNAs and bound proteins...
October 13, 2016: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/27694625/challenges-of-crispr-cas9-applications-for-long-non-coding-rna-genes
#6
Ashish Goyal, Ksenia Myacheva, Matthias Groß, Marcel Klingenberg, Berta Duran Arqué, Sven Diederichs
The CRISPR/Cas9 system provides a revolutionary genome editing tool for all areas of molecular biology. In long non-coding RNA (lncRNA) research, the Cas9 nuclease can delete lncRNA genes or introduce RNA-destabilizing elements into their locus. The nuclease-deficient dCas9 mutant retains its RNA-dependent DNA-binding activity and can modulate gene expression when fused to transcriptional repressor or activator domains. Here, we systematically analyze whether CRISPR approaches are suitable to target lncRNAs...
September 30, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27659977/capture-hybridization-analysis-of-dna-targets
#7
Alec N Sexton, Martin Machyna, Matthew D Simon
There are numerous recent cases where chromatin modifying complexes associate with long noncoding RNA (lncRNA), stoking interest in lncRNA genomic localization and associated proteins. Capture Hybridization Analysis of RNA Targets (CHART) uses complementary oligonucleotides to purify an RNA with its associated genomic DNA or proteins from formaldehyde cross-linked chromatin. Deep sequencing of the purified DNA fragments gives a comprehensive profile of the potential lncRNA biological targets in vivo. The combined identification of the genomic localization of RNA and its protein partners can directly inform hypotheses about RNA function, including recruitment of chromatin modifying complexes...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27381842/knockdown-of-gene-expression-by-antisense-morpholino-oligos-in-preimplantation-mouse-embryos-cultured-in%C3%A2-vitro
#8
Yuki Sato, Shiori Sato, Takahiro Kikuchi, Asumi Nonaka, Yuki Kumagai, Akira Sasaki, Masayuki Kobayashi
Knockdown of gene expression by antisense morpholino oligos (MOs) is a simple and effective method for analyzing the roles of genes in mammalian cells. Here, we demonstrate the efficient delivery of MOs by Endo-Porter (EP), a special transfection reagent for MOs, into preimplantation mouse embryos cultured in vitro. A fluorescein-labeled control MO was applied for monitoring the incorporation of MOs into developing 2-cell embryos in the presence of varying amounts of EP and bovine serum albumin. In optimized conditions, fluorescence was detected in 2-cell embryos within a 3-h incubation period...
September 15, 2016: Analytical Biochemistry
https://www.readbyqxmd.com/read/27328644/dazl-is-a-critical-player-for-primordial-germ-cell-formation-in-medaka
#9
Mingyou Li, Feng Zhu, Zhendong Li, Ni Hong, Yunhan Hong
The DAZ family genes boule, daz and dazl have conserved functions in primordial germ cell (PGC) migration, germ stem cell proliferation, differentiation and meiosis progression. It has remained unknown whether this family is required for PGC formation in developing embryos. Our recent study in the fish medaka (Oryzias latipes) has defined dnd as the critical PGC specifier and predicted the presence of additional factors essential for PGC formation. Here we report that dazl is a second key player for medaka PGC formation...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27285612/multi-exon-skipping-using-cocktail-antisense-oligonucleotides-in-the-canine-x-linked-muscular-dystrophy
#10
Bailey Miskew Nichols, Yoshitsugu Aoki, Mutsuki Kuraoka, Joshua J A Lee, Shin'ichi Takeda, Toshifumi Yokota
Duchenne muscular dystrophy (DMD) is one of the most common lethal genetic diseases worldwide, caused by mutations in the dystrophin (DMD) gene. Exon skipping employs short DNA/RNA-like molecules called antisense oligonucleotides (AONs) that restore the reading frame and produce shorter but functional proteins. However, exon skipping therapy faces two major hurdles: limited applicability (up to only 13% of patients can be treated with a single AON drug), and uncertain function of truncated proteins. These issues were addressed with a cocktail AON approach...
2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27197559/cyp2aa9-regulates-haematopoietic-stem-cell-development-in-zebrafish
#11
Jingying Chen, Jianbo He, Li Li, Deqin Yang, Lingfei Luo
Definitive haematopoiesis occurs during the lifetime of an individual, which continuously replenishes all blood and immune cells. During embryonic development, haematopoietic stem cell (HSC) formation is tightly controlled by growth factors, signalling molecules and transcription factors. But little is known about roles of the cytochrome P450 (CYP) 2 family member in the haematopoiesis. Here we report characterization and functional studies of Cyp2aa9, a novel zebrafish Cyp2 family member. And demonstrate that the cyp2aa9 is required for the HSC formation and homeostasis...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27141526/data-in-support-of-transcriptional-regulation-and-function-of-fas-antisense-long-noncoding-rna-during-human-erythropoiesis
#12
Olga Villamizar, Christopher B Chambers, Yin-Yuan Mo, Donald S Torry, Reese Hofstrand, Janice M Riberdy, Derek A Persons, Andrew Wilber
This paper describes data related to a research article titled, "Fas-antisense long noncoding RNA is differentially expressed during maturation of human erythrocytes and confers resistance to Fas-mediated cell death" [1]. Long noncoding RNAs (lncRNAs) are increasingly appreciated for their capacity to regulate many steps of gene expression. While recent studies suggest that many lncRNAs are functional, the scope of their actions throughout human biology is largely undefined including human red blood cell development (erythropoiesis)...
June 2016: Data in Brief
https://www.readbyqxmd.com/read/26900977/the-role-of-helper-lipids-in-lipid-nanoparticles-lnps-designed-for-oligonucleotide-delivery
#13
REVIEW
Xinwei Cheng, Robert J Lee
Lipid nanoparticles (LNPs) have shown promise as delivery vehicles for therapeutic oligonucleotides, including antisense oligos (ONs), siRNA, and microRNA mimics and inhibitors. In addition to a cationic lipid, LNPs are typically composed of helper lipids that contribute to their stability and delivery efficiency. Helper lipids with cone-shape geometry favoring the formation hexagonal II phase, such as dioleoylphosphatidylethanolamine (DOPE), can promote endosomal release of ONs. Meanwhile, cylindrical-shaped lipid phosphatidylcholine can provide greater bilayer stability, which is important for in vivo application of LNPs...
April 1, 2016: Advanced Drug Delivery Reviews
https://www.readbyqxmd.com/read/26879754/up-regulation-of-microrna-183-promotes-cell-proliferation-and-invasion-in-glioma-by-directly-targeting-nefl
#14
Ze-You Wang, Jing Xiong, Shan-Shan Zhang, Jian-Jun Wang, Zhao-Jian Gong, Min-Hui Dai
Glioblastoma multiforme (GBM) is the most common and lethal type of primary malignant brain tumor. In recent years, increasing reports suggest that discovery of microRNAs (miRNAs) might provide a novel therapeutical target for human cancers, including GBM. The expression and roles of microRNA-183 (miR-183) has been explored in several types of human cancers, including in GBM, and plays important roles in tumor initiation and progression. However, its biological functions in GBM remain largely unknown. In this study, we demonstrated that miR-183 was significantly up-regulated in astrocytoma tissues and glioblastoma cell lines...
February 15, 2016: Cellular and Molecular Neurobiology
https://www.readbyqxmd.com/read/26864466/in-vivo-knockdown-of-basal-forebrain-p75-neurotrophin-receptor-stimulates-choline-acetyltransferase-activity-in-the-mature-hippocampus
#15
Graham L Barrett, Timur Naim, Jennifer Trieu, Mengjie Huang
This study seeks to determine whether knockdown of basal forebrain p75 neurotrophin receptor (p75(NTR) ) expression elicits increased hippocampal choline acetyltransferase (ChAT) activity in mature animals. Antisense (AS) oligonucleotides (oligos) targeting p75(NTR) were infused into the medial septal area of mature rats continuously for 4 weeks. In all rats, the cannula outlet was placed equidistant between the left and the right sides of the vertical diagonal band of Broca. We tested phosphorothioate (PS), morpholino (Mo), and gapmer (mixed PS/RNA) oligos...
May 2016: Journal of Neuroscience Research
https://www.readbyqxmd.com/read/26769567/antisense-transcription-as-a-tool-to-tune-gene-expression
#16
Jennifer A N Brophy, Christopher A Voigt
A surprise that has emerged from transcriptomics is the prevalence of genomic antisense transcription, which occurs counter to gene orientation. While frequent, the roles of antisense transcription in regulation are poorly understood. We built a synthetic system in Escherichia coli to study how antisense transcription can change the expression of a gene and tune the response characteristics of a regulatory circuit. We developed a new genetic part that consists of a unidirectional terminator followed by a constitutive antisense promoter and demonstrate that this part represses gene expression proportionally to the antisense promoter strength...
January 14, 2016: Molecular Systems Biology
https://www.readbyqxmd.com/read/26680159/morpholino-antisense-oligo-inhibits-trans-splicing-of-pre-inositol-1-4-5-trisphosphate-receptor-mrna-of-trypanosoma-cruzi-and-suppresses-parasite-growth-and-infectivity
#17
Muneaki Hashimoto, Takeshi Nara, Toshihiro Mita, Katsuhiko Mikoshiba
Morpholino antisense oligos (MAOs) are used to investigate physiological gene function by inhibiting gene translation or construction of specific alternative splicing variants by blocking cis-splicing. MAOs are attractive drug candidates for viral- and bacterial-infectious disease therapy because of properties such as in vivo stability and specificity to target genes. Recently, we showed that phosphorothioate antisense oligos against Trypanosoma cruzi inositol 1,4,5-trisphosphate receptor (TcIP(3)R) mRNA inhibit the parasite host cell infection...
June 2016: Parasitology International
https://www.readbyqxmd.com/read/26530911/imaging-functional-nucleic-acid-delivery-to-skin
#18
Roger L Kaspar, Robyn P Hickerson, Emilio González-González, Manuel A Flores, Tycho P Speaker, Faye A Rogers, Leonard M Milstone, Christopher H Contag
Monogenic skin diseases arise from well-defined single gene mutations, and in some cases a single point mutation. As the target cells are superficial, these diseases are ideally suited for treatment by nucleic acid-based therapies as well as monitoring through a variety of noninvasive imaging technologies. Despite the accessibility of the skin, there remain formidable barriers for functional delivery of nucleic acids to the target cells within the dermis and epidermis. These barriers include the stratum corneum and the layered structure of the skin, as well as more locally, the cellular, endosomal and nuclear membranes...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/26464461/alu-element-containing-rnas-maintain-nucleolar-structure-and-function
#19
Maïwen Caudron-Herger, Teresa Pankert, Jeanette Seiler, Attila Németh, Renate Voit, Ingrid Grummt, Karsten Rippe
Non-coding RNAs play a key role in organizing the nucleus into functional subcompartments. By combining fluorescence microscopy and RNA deep-sequencing-based analysis, we found that RNA polymerase II transcripts originating from intronic Alu elements (aluRNAs) were enriched in the nucleolus. Antisense-oligo-mediated depletion of aluRNAs or drug-induced inhibition of RNA polymerase II activity disrupted nucleolar structure and impaired RNA polymerase I-dependent transcription of rRNA genes. In contrast, overexpression of a prototypic aluRNA sequence increased both nucleolus size and levels of pre-rRNA, suggesting a functional link between aluRNA, nucleolus integrity and pre-rRNA synthesis...
November 12, 2015: EMBO Journal
https://www.readbyqxmd.com/read/26344136/nf2-merlin-is-required-for-the-axial-pattern-formation-in-the-xenopus-laevis-embryo
#20
Xuechen Zhu, Zheying Min, Renbo Tan, Qinghua Tao
The NF2 gene product Merlin is a FERM-domain protein possessing a broad tumor-suppressing function. NF2/Merlin has been implicated in regulating multiple signaling pathways critical for cell growth and survival. However, it remains unknown whether NF2/Merlin regulates Wnt/β-catenin signaling during vertebrate embryogenesis. Here we demonstrate that NF2/Merlin is required for body pattern formation in the Xenopus laevis embryo. Depletion of the maternal NF2/Merlin enhances organizer gene expression dependent on the presence of β-catenin, and causes dorsanteriorized development; Morpholino antisense oligo-mediated knockdown of the zygotic NF2/Merlin shifts posterior genes anteriorwards and reduces the anterior development...
November 2015: Mechanisms of Development
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