primordial-germ-cell-transplantation-for-crisprcas9-based

Primordial Germ Cell Transplantation for CRISPR/Cas9-based Leapfrogging in <em>Xenopus</em>

The creation of mutant lines by genome editing is accelerating genetic analysis in many organisms. CRISPR/Cas9 methods have been adapted for use in the ...

University of California, Irvine

Diffusible morphogen models have been used widely to explain regional specification of tissues and body axes during animal development. The three-signal model for patterning the dorsal-ventral axis of the amphibian embryo proposes, in part, that a factor(s) secreted from Spemann's organizer is responsible for converting lateral marginal zone into more dorsal cell fates. We examine the possibility that chordin, a secreted inhibitor of bone morphogenetic protein (BMP) signaling and candidate "dorsalizing signal," is a long-range-acting factor. We show that chordin can, when overexpressed, act directly over distances of at least 450 microm in the early Xenopus embryo to create a gradient of BMP signaling. However, since lower levels of chordin can still induce secondary axes and these amounts of chordin act only locally to inhibit a BMP target gene, we suggest that chordin likely acts as a short-range signal in vivo. Furthermore, BMP1, a secreted metalloprotease that cleaves chordin protein in vitro, inhibits chordin's axis-inducing effects, suggesting that BMP1 functions to negatively regulate chordin's action in vivo. A dominant-negative mutant BMP1 blocks the in vitro cleavage of chordin protein by wild-type BMP1 and induces secondary axes when injected ventrally. We argue that BMP1 and Xolloid are probably functionally redundant metalloproteases and may have two roles in the early Xenopus embryo. One role may be to inhibit the action of low-level chordin protein expressed throughout the entire embryo and a possible second role may be to inhibit activation of a juxtacrine cell relay, thereby confining chordin's action to the organizer region preventing chordin from functioning as a long-range-acting factor.

Is chordin a long-range- or short-range-acting factor? Roles for BMP1-related metalloproteases in chordin and BMP4 autofeedback loop regulation.

&#x4FE1;&#x53F7;&#x901A;&#x8DEF;&#x9AA8;&#x5F62;&#x6001;&#x53D1;&#x751F;&#x86CB;&#x767D; (BMP) &#x662F;&#x5F71;&#x54CD;&#x5927;&#x5404;&#x79CD;&#x7EC6;&#x80DE;&#x884C;&#x4E3A;&#x5B88;&#x6052;&#x548C;&#x8FDB;&#x5316;&#x53E4;&#x4EE3;&#x7BA1;&#x7406;&#x6A21;&#x5757;&#x3002;&#x5927;&#x6982;&#x4F7F;&#x7528; BMP &#x54CD;&#x5E94;&#x7684;&#x8FDB;&#x5316;&#x7075;&#x6D3B;&#x6027;&#x4EA7;&#x751F;&#x7684;&#x589E;&#x9009;&#x89C4;&#x8303; BMP &#x4FE1;&#x53F7;&#x7EA7;&#x8054;&#x6765;&#x8C03;&#x8282;&#x4E0D;&#x540C;&#x7535;&#x6C60;&#x7684;&#x9776;&#x57FA;&#x56E0;&#x7684;&#x8F6C;&#x5F55;&#x3002;&#x8FD9;&#x56DE;&#x907F;&#x95EE;&#x9898;&#x7684;&#x53EF;&#x4E92;&#x6362; BMP &#x4FE1;&#x53F7;&#x4F20;&#x8F93;&#x7EC4;&#x4EF6;&#x5982;&#x4F55;&#x770B;&#x4F3C;&#x5F81;&#x6C42;&#x5E7F;&#x6CDB;&#x4E0D;&#x540C;&#x7684;&#x4EA7;&#x51FA;&#xFF0C;&#x5728;&#x4E0D;&#x540C;&#x7684;&#x5355;&#x5143;&#x683C;&#x7C7B;&#x578B;&#xFF0C;&#x4E86;&#x89E3;&#x5982;&#x4F55; BMP &#x4FE1;&#x53F7;&#x6765;&#x63A7;&#x5236;&#x53D1;&#x5C55;&#x7684;&#x57FA;&#x56E0;&#x8C03;&#x63A7;&#x7F51;&#x7EDC;&#x4E0E;&#x96C6;&#x6210;&#x7684;&#x4E0A;&#x4E0B;&#x6587;&#x4E2D;&#x7684;&#x4E00;&#x4E2A;&#x91CD;&#x8981;&#x95EE;&#x9898;&#x3002;&#x56E0;&#x4E3A;&#x5206;&#x5B50;&#x5982;&#x4F55; BMP &#x4FE1;&#x53F7;&#x6FC0;&#x6D3B;&#x9776;&#x57FA;&#x56E0;&#x7684;&#x4E0D;&#x540C;&#x7535;&#x6C60;&#x7684;&#x7406;&#x89E3;&#x662F;&#x7406;&#x89E3; BMPs &#x7684;&#x4F5C;&#x7528;&#x5728;&#x8C03;&#x8282;&#x7EC6;&#x80DE;&#x53CD;&#x5E94;&#x7684;&#x5FC5;&#x8981;&#x5148;&#x51B3;&#x6761;&#x4EF6;&#xFF0C;&#x5728;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x68C0;&#x8BA8;&#x57FA;&#x56E0;&#x9009;&#x4E2D;&#x7684;&#x4FE1;&#x53F7;&#x8F6C;&#x5BFC;&#x673A;&#x5236;&#x5982;&#x4F55; BMP &#x76D1;&#x7BA1;&#x7684;&#x76EE;&#x6807;&#x7684;&#x5F53;&#x524D;&#x7684;&#x77E5;&#x8BC6;&#x3002;&#x6211;&#x4EEC;&#x5F3A;&#x8C03;&#x6700;&#x8FD1;&#x6709;&#x7814;&#x7A76;&#x8868;&#x660E; BMP &#x76EE;&#x6807;&#x57FA;&#x56E0;&#x8C03;&#x63A7;&#x4FE1;&#x53F7;&#x7531; Schnurri &#x5BB6;&#x5EAD;&#x950C;&#x6307;&#x86CB;&#x767D;&#x7684;&#x8FDB;&#x5316;&#x517B;&#x62A4;&#x3002;&#x53D1;&#x80B2;&#x52A8;&#x6001; 238:1321-&#x7B2C; 1331&#x5E74;&#xFF0C;2009&#x5E74;&#x3002;(c) 2009&#x5E74;&#x5A01;&#x5229;&#x65AF;&#x516C;&#x53F8;&#x3002;

&#x5BFB;&#x627E;&#x5408;&#x4F5C;&#x4F19;&#x4F34;&#xFF1A; BMPs &#x662F;&#x5982;&#x4F55;&#x9009;&#x62E9;&#x4ED6;&#x4EEC;&#x7684;&#x76EE;&#x6807;&#x3002;

The bone morphogenetic protein (BMP) signaling pathway is a conserved and evolutionarily ancient regulatory module affecting a large variety of cellular behaviors. The evolutionary flexibility in using BMP responses presumably arose by co-option of a canonical BMP signaling cascade to regulate the transcription of diverse batteries of target genes. This begs the question of how seemingly interchangeable BMP signaling components elicit widely different outputs in different cell types, an important issue in the context of understanding how BMP signaling integrates with gene regulatory networks to control development. Because a molecular understanding of how BMP signaling activates different batteries of target genes is an essential prerequisite to comprehending the roles of BMPs in regulating cellular responses, here we review the current knowledge of how BMP-regulated target genes are selected by the signal transduction machinery. We highlight recent studies suggesting the evolutionary conservation of BMP target gene regulation signaling by Schnurri family zinc finger proteins. Developmental Dynamics 238:1321-1331, 2009. (c) 2009 Wiley-Liss, Inc.

Finding partners: how BMPs select their targets.

La prot&#xE9;ine morphog&#xE9;n&#xE9;tique osseuse (BMP) la voie de signalisation est un module conserv&#xE9; et de l&#x26;#39;&#xE9;volution ancienne r&#xE9;glementation touchant une grande vari&#xE9;t&#xE9; de comportements cellulaires. La flexibilit&#xE9; de l&#x26;#39;&#xE9;volution dans l&#x26;#39;utilisation de r&#xE9;ponses BMP sans doute surgi par cooptation d&#x26;#39;un canonique en cascade de signalisation BMP pour r&#xE9;guler la transcription de batteries diverses de g&#xE8;nes cibles. Cela soul&#xE8;ve la question de savoir comment apparemment interchangeables BMP composants de signalisation susciter sorties tr&#xE8;s diff&#xE9;rentes dans diff&#xE9;rents types cellulaires, une question importante dans le contexte de comprendre comment signalisation BMP int&#xE8;gre avec les r&#xE9;seaux de r&#xE9;gulation g&#xE9;n&#xE9;tique pour contr&#xF4;ler le d&#xE9;veloppement. Parce que une compr&#xE9;hension mol&#xE9;culaire de la fa&#xE7;on dont BMP signalisation active diff&#xE9;rentes batteries de g&#xE8;nes cibles est une condition essentielle &#xE0; la compr&#xE9;hension des r&#xF4;les des PGB dans la r&#xE9;gulation des r&#xE9;ponses cellulaires, ici nous passons en revue les connaissances actuelles de la fa&#xE7;on dont BMP-r&#xE9;glement&#xE9; g&#xE8;nes cibles sont s&#xE9;lectionn&#xE9;s par la machinerie de transduction du signal. Nous mettons en &#xE9;vidence des &#xE9;tudes r&#xE9;centes sugg&#xE9;rant la conservation de l&#x26;#39;&#xE9;volution de la r&#xE9;glementation BMP g&#xE8;ne cible la signalisation par les prot&#xE9;ines de la famille Schnurri doigt de zinc. Dynamique de d&#xE9;veloppement 238:1321-1331, 2009. (C) 2009 Wiley-Liss,

Recherche de partenaires: Comment MPG choisissent leurs victimes

Das Knochen morphogenetische Protein (BMP) signalisieren Weg ist Modul ein konservierten und evolution&auml;r alten ordnungspolitischen Auswirkungen auf eine Vielzahl von zellul&auml;ren Verhaltensweisen. Die evolution&auml;re Flexibilit&auml;t im Umgang mit BMP Antworten vermutlich entstand durch Kooptation von einer kanonischen BMP Signalisierung Kaskade, die Transkription von diversen Batterien Zielgene zu regulieren. Das wirft die Frage der austauschbare Komponenten der BMP-Signalisierung entlocken wie scheinbar sehr unterschiedliche Ausg&auml;nge in verschiedenen Zelltypen, ein wichtiges Thema im Zusammenhang verstehen, wie das BMP Signalisierung mit Gen regulatorische Netzwerke zur Entwicklung Steuerung integriert. Da ein molekularen Verst&auml;ndnis der wie BMP signalisieren verschiedene Batterien Zielgene aktiviert eine wesentliche Voraussetzung f&uuml;r die Rollen von BMPs in regulieren zellul&auml;re Reaktionen begreifen ist, &uuml;berpr&uuml;fen hier wir das aktuelle Wissen wie BMP-geregelt-Target, die Gene der Signal-Transduktion-Maschine ausgew&auml;hlt werden. Wir betonen neuere Studien vorgeschlagen die evolution&auml;re Erhaltung der BMP Ziel Genregulation Signalisierung durch Schnurri Familie Zink-Finger-Proteinen. Entwicklungsbiologie Dynamics 238:1321-1331, 2009. (c) 2009 Wiley-Liss, Inc.

Partnersuche: wie BMPs ihre Ziele ausw&auml;hlen.

Il bone morphogenetic protein (BMP) signaling pathway &egrave; un modulo di regolamentazione conservato ed evolutivamente antico che colpisce una grande variet&agrave; di comportamenti cellulari. La flessibilit&agrave; evolutiva utilizzando risposte BMP presumibilmente sorto di cooptazione di un canonico BMP cascata di segnalazione per regolare la trascrizione di diverse batterie di geni bersaglio. Questo solleva la questione di come apparentemente intercambiabile componenti segnalazione BMP suscitano ampiamente differenti uscite in diversi tipi di cellule, un problema importante nel contesto di comprensione come segnalazione BMP integra con reti di regolazione genica per controllare lo sviluppo. Perch&eacute; una comprensione molecolare di come BMP segnalazione attiva diverse batterie di geni bersaglio &egrave; un prerequisito essenziale per comprendere i ruoli di BMPs nel regolare le risposte cellulari, qui passiamo in rassegna le conoscenze attuali di destinazione come BMP-regolato geni sono selezionati da macchinari di trasduzione del segnale. Si evidenziano gli studi recenti suggeriscono la conservazione evolutiva della regolazione dei geni target BMP segnalazione da proteine di Schnurri famiglia zinc finger. 238:1321 Dinamica evolutiva-1331, 2009. (c) 2009 Wiley-Liss, Inc.

Ricerca di partner: come BMP selezionare i loro obiettivi.

&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x7D4C;&#x8DEF;&#x306E;&#x9AA8;&#x5F62;&#x6210;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#xFF08;BMP&#xFF09;&#x306F;&#x3001;&#x7D30;&#x80DE;&#x306E;&#x6319;&#x52D5;&#x306E;&#x591A;&#x7A2E;&#x591A;&#x69D8;&#x306A;&#x5F71;&#x97FF;&#x3092;&#x4E0E;&#x3048;&#x308B;&#x306E;&#x4FDD;&#x5B58;&#x3068;&#x9032;&#x5316;&#x7684;&#x306B;&#x53E4;&#x4EE3;&#x306E;&#x898F;&#x5236;&#x306E;&#x30E2;&#x30B8;&#x30E5;&#x30FC;&#x30EB;&#x3067;&#x3059;&#x3002; BMP&#x5FDC;&#x7B54;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x306E;&#x9032;&#x5316;&#x306E;&#x67D4;&#x8EDF;&#x6027;&#x306F;&#x3001;&#x304A;&#x305D;&#x3089;&#x304F;&#x6A19;&#x7684;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x591A;&#x69D8;&#x306A;&#x96FB;&#x6C60;&#x306E;&#x8EE2;&#x5199;&#x3092;&#x8ABF;&#x7BC0;&#x3059;&#x308B;&#x305F;&#x3081;&#x306E;&#x6A19;&#x6E96;&#x7684;&#x306A;BMP&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x30AB;&#x30B9;&#x30B1;&#x30FC;&#x30C9;&#x306E;&#x5171;&#x540C;&#x30AA;&#x30D7;&#x30B7;&#x30E7;&#x30F3;&#x3067;&#x751F;&#x307E;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x3053;&#x308C;&#x306F;&#x3001;BMP&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x6210;&#x5206;&#x304C;&#x7570;&#x306A;&#x308B;&#x7D30;&#x80DE;&#x30BF;&#x30A4;&#x30D7;&#x3001;BMP&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x304C;&#x767A;&#x9054;&#x3092;&#x5236;&#x5FA1;&#x3059;&#x308B;&#x907A;&#x4F1D;&#x5B50;&#x8ABF;&#x7BC0;&#x30CD;&#x30C3;&#x30C8;&#x30EF;&#x30FC;&#x30AF;&#x3068;&#x7D71;&#x5408;&#x3059;&#x308B;&#x65B9;&#x6CD5;&#x3092;&#x7406;&#x89E3;&#x3059;&#x308B;&#x306E;&#x30B3;&#x30F3;&#x30C6;&#x30AD;&#x30B9;&#x30C8;&#x5185;&#x3067;&#x91CD;&#x8981;&#x306A;&#x554F;&#x984C;&#x306B;&#x3082;&#x3055;&#x307E;&#x3056;&#x307E;&#x306A;&#x51FA;&#x529B;&#x3092;&#x5F15;&#x304D;&#x51FA;&#x3059;&#x65B9;&#x6CD5;&#x3001;&#x4E00;&#x898B;&#x4E92;&#x63DB;&#x6027;&#x306E;&#x554F;&#x984C;&#x3092;&#x983C;&#x3080;&#x3002; BMP&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x306F;&#x3001;&#x6A19;&#x7684;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x7570;&#x306A;&#x308B;&#x96FB;&#x6C60;&#x3092;&#x6D3B;&#x6027;&#x5316;&#x3059;&#x308B;&#x65B9;&#x6CD5;&#x306B;&#x3064;&#x3044;&#x3066;&#x306E;&#x5206;&#x5B50;&#x7684;&#x7406;&#x89E3;&#x306F;&#x3001;&#x7D30;&#x80DE;&#x5FDC;&#x7B54;&#x306E;&#x8ABF;&#x7BC0;&#x306B;BMP&#x306E;&#x5F79;&#x5272;&#x3092;&#x7406;&#x89E3;&#x306B;&#x4E0D;&#x53EF;&#x6B20;&#x306A;&#x524D;&#x63D0;&#x6761;&#x4EF6;&#x3067;&#x3042;&#x308B;&#x305F;&#x3081;&#x3001;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x6A5F;&#x69CB;&#x306B;&#x3088;&#x3063;&#x3066;&#x9078;&#x629E;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x65B9;&#x6CD5;BMP&#x8ABF;&#x7BC0;&#x6A19;&#x7684;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x73FE;&#x5728;&#x306E;&#x77E5;&#x8B58;&#x3092;&#x78BA;&#x8A8D;&#x3057;&#x307E;&#x3059;&#x3002;&#x6211;&#x3005;&#x306F;&#x3001;Schnurri&#x30D5;&#x30A1;&#x30DF;&#x30EA;&#x30FC;&#x306E;&#x30B8;&#x30F3;&#x30AF;&#x30D5;&#x30A3;&#x30F3;&#x30AC;&#x30FC;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x306B;&#x3088;&#x308B;&#x30B7;&#x30B0;&#x200B;&#x200B;&#x30CA;&#x30EB;BMP&#x306E;&#x6A19;&#x7684;&#x907A;&#x4F1D;&#x5B50;&#x767A;&#x73FE;&#x5236;&#x5FA1;&#x306E;&#x9032;&#x5316;&#x7684;&#x4FDD;&#x5B58;&#x6027;&#x3092;&#x793A;&#x5506;&#x3059;&#x308B;&#x6700;&#x8FD1;&#x306E;&#x7814;&#x7A76;&#x3092;&#x5F37;&#x8ABF;&#x8868;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x767A;&#x9054;&#x30C0;&#x30A4;&#x30CA;&#x30DF;&#x30AF;&#x30B9;238:1321-1331&#x3001;2009&#x3002; &#xFF08;C&#xFF09;2009&#x30EF;&#x30A4;&#x30EA;&#x30FC;&#x30EA;&#x30B9;&#x682A;&#x5F0F;&#x4F1A;&#x793E;

BMP&#x306F;&#x3001;&#x30BF;&#x30FC;&#x30B2;&#x30C3;&#x30C8;&#x3092;&#x9078;&#x629E;&#x3059;&#x308B;&#x65B9;&#x6CD5;&#xFF1A;&#x30D1;&#x30FC;&#x30C8;&#x30CA;&#x30FC;&#x3092;&#x898B;&#x3064;&#x3051;&#x308B;

&#xBF08; &#xC804;&#xCCB4;&#xC801; &#xB2E8;&#xBC31;&#xC9C8; (BMP) &#xC2E0;&#xD638; &#xD1B5;&#xB85C; &#xB2E4;&#xC591; &#xD55C; &#xC138;&#xD3EC; &#xD589;&#xB3D9;&#xC5D0; &#xC601;&#xD5A5;&#xC744; &#xBBF8;&#xCE58;&#xB294; &#xBCF4;&#xC874; &#xD558; &#xACE0; &#xC9C4;&#xD654; &#xC801;&#xC73C;&#xB85C; &#xACE0; &#xB300; &#xADDC;&#xC81C; &#xBAA8;&#xB4C8;. &#xC544;&#xB9C8;&#xB3C4; BMP &#xC751;&#xB2F5;&#xC744; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xC9C4;&#xD654; &#xC720;&#xC5F0;&#xC131; &#xB2E4;&#xC591; &#xD55C; &#xBC30;&#xD130;&#xB9AC; &#xB300;&#xC0C1; &#xC720;&#xC804;&#xC790;&#xC758; &#xC804;&#xC0AC; &#xC870;&#xC808; &#xD558; &#xC815;&#xC2DD; BMP &#xC2E0;&#xD638; &#xCE90;&#xC2A4;&#xCF00;&#xC774;&#xB4DC;&#xC758; co-option&#xC5D0; &#xC758;&#xD574; &#xC77C;&#xC5B4;&#xB0AC;&#xB2E4;. &#xC774; BMP &#xC2E0;&#xD638; &#xC81C;&#xC5B4; &#xAC1C;&#xBC1C; &#xD558; &#xC9C4; &#xADDC;&#xC81C; &#xB124;&#xD2B8;&#xC6CC;&#xD06C;&#xC640; &#xD1B5;&#xD569; &#xB418;&#xB294; &#xBC29;&#xC2DD;&#xC744; &#xC774;&#xD574; &#xD558;&#xB294; &#xB9E5;&#xB77D;&#xC5D0;&#xC11C; &#xC911;&#xC694; &#xD55C; &#xC774;&#xC288;&#xC778;&#xC73C;&#xB85C; &#xAD50;&#xD658;&#xD560; &#xC218; &#xC788;&#xB294; BMP &#xC2E0;&#xD638; &#xAD6C;&#xC131; &#xC694;&#xC18C; &#xB2E4;&#xB978; &#xC138;&#xD3EC; &#xC720;&#xD615;, &#xB110;&#xB9AC; &#xB2E4;&#xB978; &#xCD9C;&#xB825; &#xC720;&#xB3C4;&#xC758; &#xBB38;&#xC81C;&#xB294; &#xB0A8; &#xC8E0;. &#xC5B4;&#xB5BB;&#xAC8C; &#xB300;&#xC0C1; &#xC720;&#xC804;&#xC790;&#xC758; &#xB2E4;&#xB978; &#xBC30;&#xD130;&#xB9AC; &#xD65C;&#xC131;&#xD654; BMP &#xC2E0;&#xD638; &#xBD84;&#xC790; &#xC774;&#xD574; &#xC870;&#xC808; &#xC138;&#xD3EC;&#xC9C8; &#xC751;&#xB2F5;&#xC5D0; Bmp&#xC758; &#xC5ED;&#xD560;&#xC744; &#xC774;&#xD574; &#xD558;&#xB294; &#xD544;&#xC218;&#xC801;&#xC778; &#xC804;&#xC81C; &#xC870;&#xAC74; &#xC774;&#xBBC0;&#xB85C; &#xC5EC;&#xAE30; &#xC5B4;&#xB5BB;&#xAC8C; BMP &#xADDC;&#xC81C; &#xB300;&#xC0C1; &#xC720;&#xC804;&#xC790; &#xC2E0;&#xD638; &#xC804;&#xB2EC; &#xAE30;&#xACC4;&#xC5D0; &#xC758;&#xD574; &#xC120;&#xD0DD;&#xC758; &#xD604;&#xC7AC; &#xC815;&#xBCF4;&#xB97C; &#xAC80;&#xD1A0; &#xD569;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; &#xCD5C;&#xADFC; &#xC5F0;&#xAD6C; &#xC81C;&#xC548; BMP &#xB300;&#xC0C1; &#xC720;&#xC804;&#xC790; &#xC870;&#xC808; Schnurri &#xAC00;&#xC871; &#xC544;&#xC5F0; &#xC190;&#xAC00;&#xB77D; &#xB2E8;&#xBC31;&#xC9C8;&#xC5D0; &#xC758;&#xD574; &#xC2DC;&#xADF8;&#xB110;&#xB9C1;&#xC758; &#xC9C4;&#xD654; &#xBCF4;&#xC874;&#xC744; &#xAC15;&#xC870;. &#xBC1C;&#xB2EC; &#xC5ED;&#xD559; 238:1321-1331, 2009. (c) 2009&#xC640; &#xC77C;&#xB9AC; Liss, inc.

&#xD30C;&#xD2B8;&#xB108;&#xB97C; &#xCC3E;&#xB294;: Bmp &#xADF8;&#xB4E4;&#xC758; &#xBAA9;&#xD45C;&#xB97C; &#xC120;&#xD0DD; &#xD558;&#xB294; &#xBC29;&#xBC95;.

La prote&#xED;na morfogen&#xE9;tica &#xF3;sea (BMP) la v&#xED;a de se&#xF1;alizaci&#xF3;n es un m&#xF3;dulo de regulaci&#xF3;n conservadas evolutivamente antigua y que afecta a una gran variedad de comportamientos celulares. La flexibilidad de la evoluci&#xF3;n en el uso de las respuestas de BMP, presumiblemente surgi&#xF3; por la co-opci&#xF3;n de una can&#xF3;nica cascada de se&#xF1;alizaci&#xF3;n de BMP para regular la transcripci&#xF3;n de las bater&#xED;as de diversos de los genes diana. Esto plantea la pregunta de c&#xF3;mo aparentemente intercambiables BMP componentes de se&#xF1;alizaci&#xF3;n obtener resultados muy diferentes en diferentes tipos de c&#xE9;lulas, un tema importante en el contexto de la comprensi&#xF3;n de c&#xF3;mo la se&#xF1;alizaci&#xF3;n de BMP se integra con las redes reguladoras de genes para controlar el desarrollo. Debido a que un entendimiento molecular de c&#xF3;mo se activa la se&#xF1;alizaci&#xF3;n de BMP diferentes tipos de pilas de los genes diana es un requisito previo esencial para la comprensi&#xF3;n de los roles de las BMP en la regulaci&#xF3;n de las respuestas celulares, aqu&#xED; se revisa el conocimiento actual de c&#xF3;mo BMP-regulado genes diana son seleccionados por el mecanismo de transducci&#xF3;n de se&#xF1;ales. Destacamos los estudios recientes que sugieren que la conservaci&#xF3;n de la evoluci&#xF3;n de la regulaci&#xF3;n de BMP gen diana de se&#xF1;alizaci&#xF3;n por prote&#xED;nas dedo de zinc Schnurri familia. Din&#xE1;mica del desarrollo 238:1321-1331, 2009. (C) 2009 Wiley-Liss, Inc.

La b&#xFA;squeda de socios: &#xBF;C&#xF3;mo BMP elegir sus objetivos

The frog Xenopus laevis has for more than 60 years served as a model system for the study of vertebrate embryogenesis, molecular and cell biology, and physiology. Recently, there has been great interest in the related species Xenopus tropicalis, in part because it is diploid, unlike the allotetraploid X. laevis, and therefore amenable to forward genetics, adding to the strengths of the frog system. The genome sequence of X. tropicalis was published in 2010 and this resource is facilitating rapid progress in applying transcriptomic, genomic, proteomic, and systems biological approaches for which the Xenopus system is well suited. However, the availability of the primary nucleotide sequence is only the first step in using the genome. Accessing information embedded in the genome sequence requires well-annotated genes and knowledge of how to navigate the data. The current chapter provides a step-by-step guide for the novice to finding genes of interest in the current genome assembly, supplemented with detailed notes to improve understanding. Several publically available internet-based tools for examining the X. tropicalis genome are discussed, with special emphasis placed on the examination of synteny. Accurate determination of gene identity is enhanced by examination of orthology relationships with other organisms and thus synteny is a powerful tool. This chapter provides an access path into the Xenopus genome to enhance the researchers' ability to manipulate the organism.

Navigating the Xenopus tropicalis genome.

CRISPR/Cas9 genome editing is revolutionizing genetic loss-of-function analysis but technical limitations remain that slow progress when creating mutant lines. First, in conventional genetic breeding schemes, mosaic founder animals carrying mutant alleles are outcrossed to produce F1 heterozygotes. Phenotypic analysis occurs in the F2 generation following F1 intercrosses. Thus, mutant analyses will require multi-generational studies. Second, when targeting essential genes, efficient mutagenesis of founders is often lethal, preventing the acquisition of mature animals. Reducing mutagenesis levels may improve founder survival, but results in lower, more variable rates of germline transmission. Therefore, an efficient approach to study lethal mutations would be useful. To overcome these shortfalls, we introduce 'leapfrogging', a method combining efficient CRISPR mutagenesis with transplantation of mutated primordial germ cells into a wild-type host. Tested using Xenopus tropicalis, we show that founders containing transplants transmit mutant alleles with high efficiency. F1 offspring from intercrosses between F0 animals that carry embryonic lethal alleles recapitulate loss-of-function phenotypes, circumventing an entire generation of breeding. We anticipate that leapfrogging will be transferable to other species.

Leapfrogging: primordial germ cell transplantation permits recovery of CRISPR/Cas9-induced mutations in essential genes.

Gene regulatory networks (GRNs) involve highly combinatorial interactions between transcription factors and short sequence motifs in cis-regulatory modules of target genes to control cellular phenotypes. The GRNs specifying most cell types are largely unknown and are the subject of wide interest. A catalog of transcription factors is a valuable tool toward obtaining a deeper understanding of the role of these critical effectors in any biological setting. Here we present a comprehensive catalog of the transcription factors for the diploid frog Xenopus tropicalis. We identify 1235 genes encoding DNA-binding transcription factors, comparable to the numbers found in typical mammalian species. In detail, the repertoire of X. tropicalis transcription factor genes is nearly identical to human and mouse, with the exception of zinc finger family members, and a small number of species/lineage-specific gene duplications and losses relative to the mammalian repertoires. We applied this resource to the identification of transcription factors differentially expressed in the early gastrula stage embryo. We find transcription factor enrichment in Spemann's organizer, the ventral mesoderm, ectoderm and endoderm, and report 218 TFs that show regionalized expression patterns at this stage. Many of these have not been previously reported as expressed in the early embryo, suggesting thus far unappreciated roles for many transcription factors in the GRNs regulating early development. We expect our transcription factor catalog will facilitate myriad studies using Xenopus as a model system to understand basic biology and human disease.

Ira L. Blitz

Department of Developmental and Cell Biology

University of California, Irvine

Primordial Germ Cell Transplantation for CRISPR/Cas9-based Leapfrogging in Xenopus

Ira L. Blitz

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Developmental and Cell Biology

University of California, Irvine

Primordial Germ Cell Transplantation for CRISPR/Cas9-based Leapfrogging in Xenopus

Department of Developmental and Cell Biology