Puromycin Hydrochloride

Puromycin hydrochloride is the water-soluble hydrochloride salt of puromycin, an aminonucleoside antibiotic that blocks protein synthesis and is widely used in biomedical research. Its structure mimics the aminoacyl end of transfer RNA, allowing it to enter the ribosome’s aminoacyl-tRNA site and become incorporated into growing polypeptides, which causes premature chain termination and rapid cell death. In medicine-related research, puromycin hydrochloride is primarily used to select genetically modified mammalian cells carrying puromycin-resistance genes, verify gene delivery, and investigate translational control and cellular responses to proteotoxic stress. Its precise, potent action makes it valuable for cell biology, drug studies, and model development.

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JoVE Journal - Medicine

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Cited by 6 •

2017

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Cited by 4 •

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2026

Source: Kubala, M. H. & DeClerck, Y. A. Conditional Knockdown of Gene Expression in Cancer Cell Lines to Study the Recruitment of Monocytes/Macrophages to the Tumor Microenvironment. J. Vis. Exp. (2017)This video demonstrates the generation of conditional knockdown cancer cell lines using a Tet-inducible lentiviral shRNA system. It outlines the steps for viral transduction, antibiotic selection, and doxycycline-induced gene silencing.

Research

JoVE Journal - Cancer Research
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2026

Here, we present a protocol for generating chemoresistant cells using a dose-escalation approach, followed by a CRISPR/Cas9-based screen using a focused sgRNA library targeting epigenetic modifiers to identify regulators of acquired chemoresistance. The protocol also provides multiple optimization points tailored to chemoresistant cell models, offering a robust framework for researchers investigating resistance mechanisms.

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2025

The video outlines a process for creating genetically modified CAR T cells through the CRISPR-Cas9 System. Infecting T cells with CRISPR and CAR lentiviruses results in modifications to the target gene and the synthesis of a chimeric antigen receptor or CAR, ultimately leading to the formation of genetically modified CAR T cells.

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