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10.3389/fimmu.2021.655122

http://scihub22266oqcxt.onion/10.3389/fimmu.2021.655122
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suck abstract from ncbi


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pmid34408743      Front+Immunol 2021 ; 12 (ä): 655122
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  • Fast and Efficient Genome Editing of Human FOXP3(+) Regulatory T Cells #MMPMID34408743
  • Van Zeebroeck L; Arroyo Hornero R; Corte-Real BF; Hamad I; Meissner TB; Kleinewietfeld M
  • Front Immunol 2021[]; 12 (ä): 655122 PMID34408743show ga
  • FOXP3(+) regulatory T cells (Tregs) are central for maintaining peripheral tolerance and immune homeostasis. Because of their immunosuppressive characteristics, Tregs are a potential therapeutic target in various diseases such as autoimmunity, transplantation and infectious diseases like COVID-19. Numerous studies are currently exploring the potential of adoptive Treg therapy in different disease settings and novel genome editing techniques like CRISPR/Cas will likely widen possibilities to strengthen its efficacy. However, robust and expeditious protocols for genome editing of human Tregs are limited. Here, we describe a rapid and effective protocol for reaching high genome editing efficiencies in human Tregs without compromising cell integrity, suitable for potential therapeutic applications. By deletion of IL2RA encoding for IL-2 receptor alpha-chain (CD25) in Tregs, we demonstrated the applicability of the method for downstream functional assays and highlighted the importance for CD25 for in vitro suppressive function of human Tregs. Moreover, deletion of IL6RA (CD126) in human Tregs elicits cytokine unresponsiveness and thus may prevent IL-6-mediated instability of Tregs, making it an attractive target to potentially boost functionality in settings of adoptive Treg therapies to contain overreaching inflammation or autoimmunity. Thus, our rapid and efficient protocol for genome editing in human Tregs may advance possibilities for Treg-based cellular therapies.
  • |Blood Buffy Coat/cytology[MESH]
  • |CRISPR-Cas Systems/genetics[MESH]
  • |Forkhead Transcription Factors/metabolism[MESH]
  • |Gene Editing/*methods[MESH]
  • |Gene Knockdown Techniques[MESH]
  • |HEK293 Cells[MESH]
  • |Healthy Volunteers[MESH]
  • |Humans[MESH]
  • |Immunotherapy, Adoptive/methods[MESH]
  • |Interleukin-2 Receptor alpha Subunit/*genetics[MESH]
  • |Primary Cell Culture[MESH]
  • |RNA, Guide, CRISPR-Cas Systems/genetics[MESH]
  • |Receptors, Interleukin-6/*genetics[MESH]
  • |T-Lymphocytes, Regulatory/*metabolism[MESH]


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