Method for degrading BCL11B protein

Abstract

The invention discloses a method for degrading BCL11B protein, which is characterized by comprising the following steps: (1) obtaining Jurkat cell population capable of realizing inducible expression of a KLF4 gene; (2) expressing the KLF4 gene by adding doxycycline to promote the Jurkat cell; (3) obtaining the Jurkat cell protein and RNA of the expressed KLF4 gene; (4) detecting the SUMO conjugation of BCL11B protein obtained in the step (3) in the Jurkat cell of the expressed KLF4 gene through Western Blot detection; (5) carrying out reverse transcription synthesis on the RNA obtained in the step (3) to obtain cDNA, detecting the expression condition of the BCL11B gene by adopting real-time quantitative PCR (polymerase chain reaction) and determining that the specific protein of the BCL11B gene is degraded. The method disclosed by the invention has the advantages that a new degradation way of specific BCL11B protein is found to partially explain the apoptosis reasons of T-ALL cells to provide theoretical guidance for leukemia treatment on one hand and provide a new train of thought for realizing human T cell transdifferentiation on the other hand.

Description

technical field [0001] The invention relates to the biotechnology field of protein degradation, in particular to a method for degrading the specific protein of the BCL11B gene. Background technique [0002] The currently known protein degradation pathway is protein SUMOylation: maintaining cellular metabolism requires maintaining protein self-renewal. In addition to synthesizing proteins according to the central dogma under the guidance of genes, there is also a set of protein degradation systems in cells to maintain the desired stability of the types and quantities of proteins in cells. The ubiquitin degradation pathway of proteins was discovered in the 1980s. In 2004, the inventors Aaron Ciechanover, Avram Hershko and Irwin Rose jointly shared the Nobel Prize in Chemistry. SUMO (Small Ubiquitin-like Modifier) ​​is a kind of ubiquitin, which can covalently modify proteins to be degraded like ubiquitin molecules. It is widely involved in various cellular events, including n...

AI Technical Summary

Problems solved by technology

Overexpression of KLF4 (Kruppel like factor4) gene in T-ALL cell lines can induce apoptosis, but the downstream pathway of KLF4 and the mechanism of apoptosis have not been elucidated in detail

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