Drug Treatment and <em>In Vivo</em> Imaging of Osteoblast-Osteoclast Interactions in a Medaka Fish Osteoporosis Model

Bone-forming osteoblasts interact with bone-resorbing osteoclasts to coordinate the turnover of bone matrix and to control skeletal homeostasis. Medaka ...

National University of Singapore

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Gene synthesis is an emerging field which has widespread implications in synthetic biology and molecular biology. The field is constantly evolving which has led to key advances in oligonucleotide synthesis and gene synthesis technologies, with simplicity, cost effectiveness and high throughput. The miniaturization, multiplexing, microfluidic processing and the integrated microchip engineering will drive down cost and increase productivity without compromising DNA synthesis fidelity, whereas the gigantic amount of genome information provides infinite source of DNA elements and genes as raw material for synthetic biology. This article describes some of the recent patents on oligonucleotide synthesis and gene synthesis.

Recent patents on oligonucleotide synthesis and gene synthesis.

Synth&egrave;se de g&egrave;ne est un domaine &eacute;mergent qui a des implications tr&egrave;s r&eacute;pandues en biologie mol&eacute;culaire et la biologie synth&eacute;tique. Le domaine est en constante &eacute;volution qui a conduit &agrave; des avances principales dans la synth&egrave;se d&#39;oligonucl&eacute;otides et synth&egrave;se de g&egrave;ne technologies, avec simplicit&eacute;, rentabilit&eacute; et haut d&eacute;bit. La miniaturisation, multiplexage, traitement de microfluidique et l&#39;ing&eacute;nierie de la micropuce int&eacute;gr&eacute;e vont baisser les co&ucirc;ts et augmenter la productivit&eacute; sans compromettre fid&eacute;lit&eacute; de synth&egrave;se de l&#39;ADN, alors que la quantit&eacute; gigantesque d&#39;informations de g&eacute;nome fournit une source infinie de g&egrave;nes et des &eacute;l&eacute;ments d&#39;ADN comme mati&egrave;re premi&egrave;re pour la biologie synth&eacute;tique. Cet article d&eacute;crit certains des brevets r&eacute;cents sur la synth&egrave;se d&#39;oligonucl&eacute;otides et synth&egrave;se de g&egrave;ne.

Brevets r&eacute;cents sur la synth&egrave;se d&#39;oligonucl&eacute;otides et synth&egrave;se de g&egrave;ne.

Gensynthese ist ein aufstrebenden Gebiet die weitverbreitete Auswirkungen in der synthetischen Biologie und Molekulare Biologie hat. Das Feld wird st&auml;ndig weiterentwickelt, die wichtige Fortschritte in der Oligonukleotid Synthese und Gensynthese mit Einfachheit, Wirtschaftlichkeit und Hochdurchsatz-Technologien gef&uuml;hrt hat. Die Miniaturisierung, multiplexing, Microfluidic Verarbeitung und das integrierte Mikrochip-Engineering werden senken Kosten und erh&ouml;hen die Produktivit&auml;t ohne Kompromisse bei DNA-Synthese-Treue, die riesige Menge von Genominformationen unendliche Quelle der DNA-Elemente und Gene als Rohstoff f&uuml;r synthetische Biologie sieht. Dieser Artikel beschreibt einige der j&uuml;ngsten Patente auf Oligonukleotid Synthese und Gensynthese.

Aktuelle Patente auf Oligonukleotid Synthese und Gensynthese.

Sintesi del gene &egrave; un settore emergente che ha implicazioni molto diffuse in biologia molecolare e biologia sintetica. Il campo &egrave; in continua evoluzione che ha portato a progressi chiave nella sintesi del oligonucleotide e gene sintesi technologies, con semplicit&agrave;, costo efficacia e produttivit&agrave; elevata. La miniaturizzazione, multiplexing, elaborazione microfluidica e l&#39;ingegneria integrata microchip sar&agrave; guidare verso il basso costo e aumentare la produttivit&agrave; senza compromettere fedelt&agrave; di sintesi del DNA, considerando la quantit&agrave; gigantesca di informazioni sul genoma fonte infinita di elementi del DNA e dei geni come materia prima per la biologia sintetica. In questo articolo vengono descritti alcuni dei brevetti recenti sulla sintesi del oligonucleotide e gene sintesi.

Recenti brevetti sulla sintesi del oligonucleotide e gene sintesi.

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&#xC720;&#xC804;&#xC790; &#xD569;&#xC131; &#xD569;&#xC131; &#xC0DD;&#xBB3C;&#xD559; &#xBC0F; &#xBD84;&#xC790; &#xC0DD;&#xBB3C;&#xD559;&#xC5D0; &#xAD11;&#xBC94;&#xC704; &#xD55C; &#xC758;&#xBBF8;&#xB97C;&#xAC00;&#xC9C0;&#xACE0; &#xC788;&#xB294; &#xC2E0;&#xD765; &#xBD84;&#xC57C; &#xC774;&#xB2E4;. &#xD544;&#xB4DC; &#xB04A;&#xC784;&#xC5C6;&#xC774; &#xBCC0;&#xD654; &#xD558;&#xB294; &#xAE30;&#xC220;, &#xB2E8;&#xC21C;, &#xBE44;&#xC6A9; &#xD6A8;&#xACFC; &#xBC0F; &#xB192;&#xC740; &#xCC98;&#xB9AC;&#xB7C9; oligonucleotide &#xD569;&#xC131; &#xBC0F; &#xC720;&#xC804;&#xC790; &#xD569;&#xC131;&#xC758; &#xC8FC;&#xC694; &#xBC1C;&#xC804;&#xC744; &#xC8FC;&#xB3C4;&#xD558; &#xACE0; &#xC788;&#xB2E4;. &#xC18C;&#xD615;&#xD654;, &#xBA40;&#xD2F0;&#xD50C;&#xB809;&#xC2F1;, &#xBBF8;&#xC138; &#xAC00;&#xACF5; &#xBC0F; &#xD1B5;&#xD569;&#xB41C; &#xB9C8;&#xC774;&#xD06C;&#xB85C;&#xCE69; &#xC5D4;&#xC9C0;&#xB2C8;&#xC5B4;&#xB9C1; &#xBE44;&#xC6A9; &#xC808;&#xAC10; &#xB418;&#xBA70; &#xAC70; &#xB300; &#xD55C; &#xC591;&#xC758; &#xAC8C;&#xB188; &#xC815;&#xBCF4; &#xD569;&#xC131; &#xC0DD;&#xBB3C;&#xD559;&#xC5D0; &#xB300; &#xD55C; &#xC6D0;&#xB8CC;&#xB85C; DNA &#xC694;&#xC18C;&#xC640; &#xC720;&#xC804;&#xC790;&#xC758; &#xBB34;&#xD55C; &#xD55C; &#xC18C;&#xC2A4;&#xB97C; &#xC81C;&#xACF5; &#xD558;&#xB294; &#xBC18;&#xBA74; &#xC190;&#xC0C1; DNA &#xD569;&#xC131; &#xCDA9;&#xC2E4;&#xB3C4; &#xC5C6;&#xC774; &#xC0DD;&#xC0B0;&#xC131;&#xC744; &#xD5A5;&#xC0C1;. &#xC774; &#xBB38;&#xC11C; oligonucleotide &#xD569;&#xC131; &#xBC0F; &#xC720;&#xC804;&#xC790; &#xD569;&#xC131;&#xC5D0; &#xCD5C;&#xADFC; &#xD2B9;&#xD5C8; &#xC911; &#xC77C;&#xBD80;&#xB97C; &#xC124;&#xBA85;&#xD569;&#xB2C8;&#xB2E4;.

Oligonucleotide &#xD569;&#xC131; &#xBC0F; &#xC720;&#xC804;&#xC790; &#xD569;&#xC131;&#xC5D0; &#xCD5C;&#xADFC; &#xD2B9;&#xD5C8;&#xC785;&#xB2C8;&#xB2E4;.

S&iacute;ntesis de genes es un campo emergente que tiene implicaciones generalizadas en biolog&iacute;a sint&eacute;tica y biolog&iacute;a molecular. El campo est&aacute; en constante evoluci&oacute;n que ha llevado a avances claves en la s&iacute;ntesis de oligonucle&oacute;tidos y s&iacute;ntesis de gene technologies, con sencillez, costo efectividad y alto rendimiento. La miniaturizaci&oacute;n, multiplexaci&oacute;n, microflu&iacute;dica Ingenier&iacute;a de proceso y el microchip integrado se disminuyan los costos y aumentar la productividad sin comprometer fidelidad de s&iacute;ntesis de ADN, mientras que la enorme cantidad de informaci&oacute;n de genoma proporciona una fuente infinita de elementos de ADN y genes como materia prima para la biolog&iacute;a sint&eacute;tica. Este art&iacute;culo describe algunas de las recientes patentes en la s&iacute;ntesis de oligonucle&oacute;tidos y s&iacute;ntesis de genes.

Recientes patentes sobre s&iacute;ntesis de oligonucle&oacute;tidos y s&iacute;ntesis de genes.

During vertebrate neurulation, cranial neural crest cells (CNCCs) undergo epithelial to mesenchymal transition (EMT), delaminate from the neural plate border, and migrate as separate streams into different cranial regions. There, they differentiate into distinct parts of the craniofacial skeleton. Canonical Wnt signaling has been shown to be essential for this process at different levels but the involved receptors remained unclear. Here we show that the frizzled co-receptor low-density-lipoprotein (LDL) receptor-related protein 5 (Lrp5) plays a crucial role in CNCC migration and morphogenesis of the cranial skeleton. Early during induction and migration of CNCCs, lrp5 is expressed ubiquitously but later gets restricted to CNCC derivatives in the ventral head region besides different regions in the CNS. A knock-down of lrp5 does not interfere with induction of CNCCs but leads to reduced proliferation of premigratory CNCCs. In addition, cell migration is disrupted as CNCCs are found in clusters at ectopic positions in the dorsomedial neuroepithelium after lrp5 knock-down and transient CRISPR/Cas9 gene editing. These migratory defects consequently result in malformations of the craniofacial skeleton. To date, Lrp5 has mainly been associated with bone homeostasis in mammals. Here we show that in zebrafish, lrp5 also controls cell migration during early morphogenetic processes and contributes to shaping the craniofacial skeleton.

The Wnt Co-Receptor Lrp5 Is Required for Cranial Neural Crest Cell Migration in Zebrafish.

Osteoclasts are bone-resorbing cells derived from the monocyte/macrophage lineage. Excess osteoclast activity leads to reduced bone mineral density, a hallmark of diseases such as osteoporosis. Processes that regulate osteoclast activity are therefore targeted in current osteoporosis therapies. To identify and characterize drugs for treatment of bone diseases, suitable in vivo models are needed to complement cell-culture assays. We have previously reported transgenic medaka lines expressing the osteoclast-inducing factor receptor activator of nuclear factor κB ligand (Rankl) under control of a heat shock-inducible promoter. Forced Rankl expression resulted in ectopic osteoclast formation, as visualized by live imaging in fluorescent reporter lines. This led to increased bone resorption and a dramatic reduction of mineralized matrix similar to the situation in humans with osteoporosis. In an attempt to establish the medaka as an in vivo model for osteoporosis drug screening, we treated Rankl-expressing larvae with etidronate and alendronate, two bisphosphonates commonly used in human osteoporosis therapy. Using live imaging, we observed an efficient, dose-dependent inhibition of osteoclast activity, which resulted in the maintenance of bone integrity despite an excess of osteoclast formation. Strikingly, we also found that bone recovery was efficiently promoted after inhibition of osteoclast activity and that osteoblast distribution was altered, suggesting effects on osteoblast-osteoclast coupling. Our data show that transgenic medaka lines are suitable in vivo models for the characterization of antiresorptive or bone-anabolic compounds by live imaging and for screening of novel osteoporosis drugs.

Live imaging of osteoclast inhibition by bisphosphonates in a medaka osteoporosis model.

Sp7/osterix (osx) encodes a zinc-finger transcription factor that controls osteoblast differentiation in mammals. Although identified in all vertebrate lineages, its role in non-mammalian bone formation remains elusive. Here we show that an osx mutation in medaka results in severe bone defects and larval lethality. Pre-osteoblasts fail to differentiate leading to severe intramembranous and perichondral ossification defects. The notochord sheath mineralizes normally supporting the idea of an osteoblast-independent mechanism for teleost vertebral centra formation. This study establishes a key role for Sp7/Osx for bone formation also in a non-mammalian species, and reveals conserved and non-conserved features in vertebrate bone formation.

Tingsheng Yu

Department of Biological Sciences,
NUS Centre for Bioimaging Sciences (CBIS)

National University of Singapore

Drug Treatment and In Vivo Imaging of Osteoblast-Osteoclast Interactions in a Medaka Fish Osteoporosis Model

Tingsheng Yu

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Biological Sciences,NUS Centre for Bioimaging Sciences (CBIS)

National University of Singapore

Drug Treatment and In Vivo Imaging of Osteoblast-Osteoclast Interactions in a Medaka Fish Osteoporosis Model

Department of Biological Sciences,
NUS Centre for Bioimaging Sciences (CBIS)