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Protocol for genomic editing in human resting primary NK cells and NK-92 cells via CRISPR-Cas9 ribonucleoproteins.
Verhezen, Tias; Lau, Ho Wa; Van Audenaerde, Jonas; Wouters, An; Smits, Evelien; De Waele, Jorrit.
Afiliação
  • Verhezen T; Center for Oncological Research, Integrated Precision and Personalized Oncology Network (IPPON), University of Antwerp, Antwerp, Belgium. Electronic address: tias.verhezen@uantwerpen.be.
  • Lau HW; Center for Oncological Research, Integrated Precision and Personalized Oncology Network (IPPON), University of Antwerp, Antwerp, Belgium.
  • Van Audenaerde J; Center for Oncological Research, Integrated Precision and Personalized Oncology Network (IPPON), University of Antwerp, Antwerp, Belgium.
  • Wouters A; Center for Oncological Research, Integrated Precision and Personalized Oncology Network (IPPON), University of Antwerp, Antwerp, Belgium.
  • Smits E; Center for Oncological Research, Integrated Precision and Personalized Oncology Network (IPPON), University of Antwerp, Antwerp, Belgium.
  • De Waele J; Center for Oncological Research, Integrated Precision and Personalized Oncology Network (IPPON), University of Antwerp, Antwerp, Belgium. Electronic address: jorrit.dewaele@uantwerpen.be.
STAR Protoc ; 5(3): 103123, 2024 Sep 20.
Article em En | MEDLINE | ID: mdl-38935511
ABSTRACT
Here, we present a protocol to perform CRISPR-Cas9 genome editing in human resting primary natural killer (NK) and NK-92 cells. We describe steps for guide RNA selection, ribonucleoprotein (RNP) complex formation, delivery via Nucleofection, and analysis of CRISPR edits to assess editing efficiencies. This protocol offers a tool for functional studies in NK cells, paving the way for potential applications in immunotherapy and beyond. We also discuss limitations such as off-target effects and cell-type-specific considerations.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Ribonucleoproteínas / Células Matadoras Naturais / Sistemas CRISPR-Cas / Edição de Genes / RNA Guia de Sistemas CRISPR-Cas Limite: Humans Idioma: En Revista: STAR Protoc Ano de publicação: 2024 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Ribonucleoproteínas / Células Matadoras Naturais / Sistemas CRISPR-Cas / Edição de Genes / RNA Guia de Sistemas CRISPR-Cas Limite: Humans Idioma: En Revista: STAR Protoc Ano de publicação: 2024 Tipo de documento: Article