Inflammasome : Investigating the effect of NEK7 in the activation of the NLRP3 Inflammasome

• Main Author: Adindu Uzowuru, Cosmas
• Format: Others
• Language: English
• Published: Högskolan i Skövde, Institutionen för biovetenskap 2020
• Subjects: NLRP3 inflammasome; NEK7 protein; THP-1; Prime; activate; differentiate; stimulate; cytokines; Interleukin-1 Beta; Interleukin-18; Medical Bioscience; Medicinsk biovetenskap
• Online Access: http://urn.kb.se/resolve?urn=urn:nbn:se:his:diva-18937

Inflammation is a biological defence mechanism applied by living organisms against foreign invaders. In the response to DAMPs and PAMPs, organisms use inflammatory multi-protein complexes to fight the attackers. The most studied inflammasome proteins are NLRP3, ASC and Caspase-1. This study is aimed at understanding the role of NEK7 protein in the NLRP3 inflammasome’s activation, using CRISPR/Cas9 system. To determine the effect of CRISPR/Cas9 and transfection, mRNA expression was analyzed. The results obtained suggest that neither the transfection nor the NEK7 protein knockout have sufficiently worked. This study could not experimentally establish that NEK7 triggers NLRP3 inflammasome activation because ELISA was not conducted to verify the levels of cytokines emitted, due to there being no statistical differences between the samples. Above all, the research question in this thesis project was not answered because the instability of the ACTB reference gene negatively influenced the results. However, previous related studies conclude that NEK7 plays a crucial role in the activation of the NLRP3 inflammasome.