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Statements

Subject Item
dbr:Cre-Lox_recombination
rdfs:label
LoxP-Cre rekombineringssystem Cre-Lox重组 Recombinação Cre-Lox Cre-Lox recombination Cre/loxP-System Cre-loxP部位特異的組換え
rdfs:comment
Cre-loxP部位特異的組換えは、1981年にバクテリオファージP1の研究で見出された反応である。loxPという特定のDNA配列を標的としており、DNA組換え酵素Creにより触媒される。1987年デュポン社（当時）のBrian Sauerが真核生物での応用法を開発したのを端緒に、現在では条件的遺伝子ノックアウトを実施する目的などで広く使われる技術となっている。 loxP-Cre rekombineringssystemet är en molekylärgenetisk metod för att undersöka effekten av olika gener i specifika vävnader. Metoden används för att skapa en så kallad knockoutmus, eller annan knockoutorganism, där en viss gen i en viss celltyp stängs av. loxP är speciella rekombineringssiter på DNA, en form av repeterade sekvenser som kan omge ett exon. Cre är en gen som kodar för ett protein som binder till IoxP-sekvenser på DNA-strängen, och inducerar homolog överkorsning över de två IoxP-fragmenten så att DNA-materialet mellan loxP-fragmenten spjälkas bort och bildar en liten plasmid. Das Cre/loxP-System ist ein Rekombinations-System. Es ermöglicht das gezielte Entfernen von DNA-Sequenzen in lebenden Organismen. Mit dieser Technik können beispielsweise einzelne spezifische Zell- oder Gewebearten genetisch modifiziert werden, während andere Gewebe davon unberührt bleiben. Ursprünglich stammt dieses System aus dem Bakteriophagen P1 und findet heute v. a. in der molekularbiologischen Forschung und zur Herstellung gentechnisch veränderter Pflanzen Anwendung. Ein analoges System ist das Flp-FRT-System. Cre-Lox recombination is a site-specific recombinase technology, used to carry out deletions, insertions, translocations and inversions at specific sites in the DNA of cells. It allows the DNA modification to be targeted to a specific cell type or be triggered by a specific external stimulus. It is implemented both in eukaryotic and prokaryotic systems. The Cre-lox recombination system has been particularly useful to help neuroscientists to study the brain in which complex cell types and neural circuits come together to generate cognition and behaviors. NIH Blueprint for Neuroscience Research has created several hundreds of Cre driver mouse lines which are currently used by the worldwide neuroscience community. Cre-Lox重组是一种用于在细胞DNA的特定位点上进行删除、插入、转座和操作的，其特点在于可以让DNA修改对指定的特定细胞群进行，或是使修改过程由特定的外部刺激所触发。此技术对真核和原核细胞都适用。特别地，对研究由各种复杂细胞和神经通路复合而形成认知行为的大脑的而言，这种技术已被证明是相当实用的。 这个重组体系由单个酶组成，即可以让一对被称为Lox序列的短目标序列进行重组的，除此之外不需要插入其他的额外蛋白或序列。Cre重组酶和源Lox序列（即LoxP序列）是从P1噬菌体中获取的。 通过合适地插入Lox序列可以使特定基因活化、沉默或者和其他基因交换位置——在DNA的层面上，许多操作都可以进行。Cre酶的活性可控，可以使之只在一种特定细胞类型中表达，也可以使之由特定的外部刺激（如化学信号、热刺激）触发表达。这些特异性DNA修饰对细胞系追踪以及有致命性的突变研究十分有效。 Cre-Lox重组系统在操作和用途上都与系统十分相似。 Recombinação Cre-Lox é uma tecnologia de recombinação sítio-específica, cujo nome vem do uso da enzima e . Derivada do mecanismo natural de recombinação do , a recombinação Cre-Lox é utilizada para diversos usos científicos e tecnológicos, notavelmente inserções, deleções, inversões e translocações sítio-específicas em sequências de DNA. Diz-se que uma sequência é “floxed” quando está flanqueada por sítios Lox.
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dbc:Molecular_genetics dbc:Genetic_engineering dbc:DNA dbc:Genetics_techniques
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4363576
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loxP-Cre rekombineringssystemet är en molekylärgenetisk metod för att undersöka effekten av olika gener i specifika vävnader. Metoden används för att skapa en så kallad knockoutmus, eller annan knockoutorganism, där en viss gen i en viss celltyp stängs av. loxP är speciella rekombineringssiter på DNA, en form av repeterade sekvenser som kan omge ett exon. Cre är en gen som kodar för ett protein som binder till IoxP-sekvenser på DNA-strängen, och inducerar homolog överkorsning över de två IoxP-fragmenten så att DNA-materialet mellan loxP-fragmenten spjälkas bort och bildar en liten plasmid. Rent praktiskt går metoden ut på att via homolog rekombination skapa möss som både har loxP-sekvenser som omger genen man vill stänga av, samt som har en gen för Cre som styrs av en celltyps-specifik promotor. När cellen differentierats till en celltyp som producerar aktivatorer som aktiverar promotorn för Cre-genen så kommer Cre-protein att bildas, som kommer att spjälka bort och därmed förstöra genen som omges av loxP-sekvens. Das Cre/loxP-System ist ein Rekombinations-System. Es ermöglicht das gezielte Entfernen von DNA-Sequenzen in lebenden Organismen. Mit dieser Technik können beispielsweise einzelne spezifische Zell- oder Gewebearten genetisch modifiziert werden, während andere Gewebe davon unberührt bleiben. Ursprünglich stammt dieses System aus dem Bakteriophagen P1 und findet heute v. a. in der molekularbiologischen Forschung und zur Herstellung gentechnisch veränderter Pflanzen Anwendung. Ein analoges System ist das Flp-FRT-System. Cre-loxP部位特異的組換えは、1981年にバクテリオファージP1の研究で見出された反応である。loxPという特定のDNA配列を標的としており、DNA組換え酵素Creにより触媒される。1987年デュポン社（当時）のBrian Sauerが真核生物での応用法を開発したのを端緒に、現在では条件的遺伝子ノックアウトを実施する目的などで広く使われる技術となっている。 Cre-Lox重组是一种用于在细胞DNA的特定位点上进行删除、插入、转座和操作的，其特点在于可以让DNA修改对指定的特定细胞群进行，或是使修改过程由特定的外部刺激所触发。此技术对真核和原核细胞都适用。特别地，对研究由各种复杂细胞和神经通路复合而形成认知行为的大脑的而言，这种技术已被证明是相当实用的。 这个重组体系由单个酶组成，即可以让一对被称为Lox序列的短目标序列进行重组的，除此之外不需要插入其他的额外蛋白或序列。Cre重组酶和源Lox序列（即LoxP序列）是从P1噬菌体中获取的。 通过合适地插入Lox序列可以使特定基因活化、沉默或者和其他基因交换位置——在DNA的层面上，许多操作都可以进行。Cre酶的活性可控，可以使之只在一种特定细胞类型中表达，也可以使之由特定的外部刺激（如化学信号、热刺激）触发表达。这些特异性DNA修饰对细胞系追踪以及有致命性的突变研究十分有效。 Cre-Lox重组系统在操作和用途上都与系统十分相似。 Cre-Lox recombination is a site-specific recombinase technology, used to carry out deletions, insertions, translocations and inversions at specific sites in the DNA of cells. It allows the DNA modification to be targeted to a specific cell type or be triggered by a specific external stimulus. It is implemented both in eukaryotic and prokaryotic systems. The Cre-lox recombination system has been particularly useful to help neuroscientists to study the brain in which complex cell types and neural circuits come together to generate cognition and behaviors. NIH Blueprint for Neuroscience Research has created several hundreds of Cre driver mouse lines which are currently used by the worldwide neuroscience community. The system consists of a single enzyme, Cre recombinase, that recombines a pair of short target sequences called the Lox sequences. This system can be implemented without inserting any extra supporting proteins or sequences. The Cre enzyme and the original Lox site called the LoxP sequence are derived from bacteriophage P1. Placing Lox sequences appropriately allows genes to be activated, repressed, or exchanged for other genes. At a DNA level many types of manipulations can be carried out. The activity of the Cre enzyme can be controlled so that it is expressed in a particular cell type or triggered by an external stimulus like a chemical signal or a heat shock. These targeted DNA changes are useful in cell lineage tracing and when mutants are lethal if expressed globally. The Cre-Lox system is very similar in action and in usage to the FLP-FRT recombination system. Recombinação Cre-Lox é uma tecnologia de recombinação sítio-específica, cujo nome vem do uso da enzima e . Derivada do mecanismo natural de recombinação do , a recombinação Cre-Lox é utilizada para diversos usos científicos e tecnológicos, notavelmente inserções, deleções, inversões e translocações sítio-específicas em sequências de DNA. Diz-se que uma sequência é “floxed” quando está flanqueada por sítios Lox. Um dos usos mais conhecidos da recombinação Cre-Lox é a criação de animais knockout condicionais, onde a modificação desejada ocorrerá apenas após um estímulo desejado. Isso permite a alteração de sequências de DNA em tecidos específicos ou em momentos específicos do desenvolvimento, driblando possíveis efeitos letais se a alteração ocorresse globalmente.
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