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CRISPRi as an efficient tool for gene repression in archaea.
Stachler, Aris-Edda; Schwarz, Thandi Sarah; Schreiber, Sandra; Marchfelder, Anita.
Afiliación
  • Stachler AE; Biology II, Ulm University, 89069 Ulm, Germany.
  • Schwarz TS; Biology II, Ulm University, 89069 Ulm, Germany.
  • Schreiber S; Biology II, Ulm University, 89069 Ulm, Germany.
  • Marchfelder A; Biology II, Ulm University, 89069 Ulm, Germany. Electronic address: anita.marchfelder@uni-ulm.de.
Methods ; 172: 76-85, 2020 02 01.
Article en En | MEDLINE | ID: mdl-31150759
In the years following its discovery and characterization, the CRISPR-Cas system has been modified and converted into a multitude of applications for eukaryotes and bacteria, such as genome editing and gene regulation. Since no such method has been available for archaea, we developed a tool for gene repression in the haloarchaeon Haloferax volcanii by repurposing its endogenous type I-B CRISPR-Cas system. Here, we present the two possible approaches for gene repression as well as our workflow to achieve and assess gene knockdown, offer recommendations on protospacer selection and give some examples of genes we have successfully silenced.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Haloferax volcanii / Regulación de la Expresión Génica Arqueal / Sistemas CRISPR-Cas / Edición Génica Idioma: En Revista: Methods Asunto de la revista: BIOQUIMICA Año: 2020 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Haloferax volcanii / Regulación de la Expresión Génica Arqueal / Sistemas CRISPR-Cas / Edición Génica Idioma: En Revista: Methods Asunto de la revista: BIOQUIMICA Año: 2020 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Estados Unidos