Description

• Background: MLKL (Mixed Lineage Kinase Domain-Like Protein) is a pseudokinase and serves as a key effector molecule in the RIPK (Receptor-Interacting Protein Kinase) signaling pathway. It mediates the execution of programmed necroptosis and plays a role in inflammation, immune regulation, and disease pathogenesis.Upon activation, MLKL oligomerizes or polymerizes and translocates to the plasma membrane, leading to membrane disruption and subsequent necrotic cell death.
• Targeting strategy: The exons 2-10 of mouse Mlkl gene were knockout in B-Mlkl KO mice. Mouse Mlkl gene transcription and translation will be disrupted.
• Validation: Mouse MLKL protein was not detectable in B-Mlkl KO mice.Mouse Mlkl mRNA was exclusively detectable in C57BL/6N but not detectable in B-Mlkl KO mice.
• Application：Pharmacological Efficacy Evaluation for Fatty Liver Disease and Hepatic Ischemia-Reperfusion Injury.

Targeting strategy

Gene targeting strategy for B-Mlkl KO mice. The exons 2-10 of mouse Mlkl gene were knockout in B-Mlkl KO mice. Mouse Mlkl gene transcription and translation will be disrupted.

Protein expression analysis

Western blot analysis of Mlkl protein expression in homozygous B-Mlkl KO mice. Various tissue lysates were collected from wild-type C57BL/6N mice (+/+) and homozygous B-Mlkl KO mice (-/-) (male, 6-week-old), and then analyzed by western blot with anti-Mlkl antibody(abcam, ab243142). 40 μg total proteins were loaded for western blotting analysis. MLKL was not detectable in liver, lung, brain, stomach and kidney of B-Mlkl KO mice.

mRNA expression analysis

Strain specific analysis of Mlkl mRNA expression in wild-type C57BL/6N and B-Mlkl KO mice by RT-PCR. Liver RNA were isolated from wild-type C57BL/6N (+/+) and homozygous B-Mlkl KO mice(-/-), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse Mlkl primers. Mouse Mlkl mRNA was exclusively detectable in C57BL/6N but not detectable in B-Mlkl KO mice.