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MELK expression correlates with tumor mitotic activity but is not required for cancer growth.

eLife | 2018

The Maternal Embryonic Leucine Zipper Kinase (MELK) has been identified as a promising therapeutic target in multiple cancer types. MELK over-expression is associated with aggressive disease, and MELK has been implicated in numerous cancer-related processes, including chemotherapy resistance, stem cell renewal, and tumor growth. Previously, we established that triple-negative breast cancer cell lines harboring CRISPR/Cas9-induced null mutations in MELK proliferate at wild-type levels in vitro (Lin et al., 2017). Here, we generate several additional knockout clones of MELK and demonstrate that across cancer types, cells lacking MELK exhibit wild-type growth in vitro, under environmental stress, in the presence of cytotoxic chemotherapies, and in vivo. By combining our MELK-knockout clones with a recently described, highly specific MELK inhibitor, we further demonstrate that the acute inhibition of MELK results in no specific anti-proliferative phenotype. Analysis of gene expression data from cohorts of cancer patients identifies MELK expression as a correlate of tumor mitotic activity, explaining its association with poor clinical prognosis. In total, our results demonstrate the power of CRISPR/Cas9-based genetic approaches to investigate cancer drug targets, and call into question the rationale for treating patients with anti-MELK monotherapies.

Pubmed ID: 29417930 RIS Download

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Addgene (tool)

RRID:SCR_002037

Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.

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Jackson Laboratory (tool)

RRID:SCR_004633

An independent, nonprofit organization focused on mammalian genetics research to advance human health. Their mission is to discover the genetic basis for preventing, treating, and curing human disease, and to enable research for the global biomedical community. Jackson Laboratory breeds and manages colonies of mice as resources for other research institutions and laboratories, along with providing software and techniques. Jackson Lab also conducts genetic research and provides educational material for various educational levels.

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Cold Spring Harbor Laboratory (tool)

RRID:SCR_008326

Non profit, private research and education institution that performs molecular and genetic research used to generate methods for better diagnostics and treatments for cancer and neurological diseases. Research of cancer causing genes and their respective signaling pathways, mutations and structural variations of the human genome that could cause neurodevelopmental and neurodegenerative illnesses such as autism, schizophrenia, and Alzheimer's and Parkinson's diseases and also research in plant genetics and quantitative biology.

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RRID:SCR_008426

Commercial instrument and chemical vendor. Developer and manufacturer of specialized technological products for life science research and clinical diagnostics markets.

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A-375 (cell line)

RRID:CVCL_0132

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DLD-1 (cell line)

RRID:CVCL_0248

Cell line DLD-1 is a Cancer cell line with a species of origin Homo sapiens (Human)

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CAL-51 (cell line)

RRID:CVCL_1110

Cell line CAL-51 is a Cancer cell line with a species of origin Homo sapiens

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Rat1A (cell line)

RRID:CVCL_0512

Cell line Rat1A is a Spontaneously immortalized cell line with a species of origin Rattus norvegicus

View all literature mentions

A-375 (cell line)

RRID:CVCL_0132

Cell line A-375 is a Cancer cell line with a species of origin Homo sapiens (Human)

View all literature mentions

MDA-MB-231 (cell line)

RRID:CVCL_0062

Cell line MDA-MB-231 is a Cancer cell line with a species of origin Homo sapiens (Human)

View all literature mentions

NIH 3T3 (cell line)

RRID:CVCL_0594

Cell line NIH 3T3 is a Spontaneously immortalized cell line with a species of origin Mus musculus

View all literature mentions

CAL-51 (cell line)

RRID:CVCL_1110

Cell line CAL-51 is a Cancer cell line with a species of origin Homo sapiens

View all literature mentions

A-375 (cell line)

RRID:CVCL_0132

Cell line A-375 is a Cancer cell line with a species of origin Homo sapiens (Human)

View all literature mentions

MCF-10A (cell line)

RRID:CVCL_0598

Cell line MCF-10A is a Spontaneously immortalized cell line with a species of origin Homo sapiens

View all literature mentions

NIH 3T3 (cell line)

RRID:CVCL_0594

Cell line NIH 3T3 is a Spontaneously immortalized cell line with a species of origin Mus musculus

View all literature mentions

MCF-10A (cell line)

RRID:CVCL_0598

Cell line MCF-10A is a Spontaneously immortalized cell line with a species of origin Homo sapiens

View all literature mentions

Rat1A (cell line)

RRID:CVCL_0512

Cell line Rat1A is a Spontaneously immortalized cell line with a species of origin Rattus norvegicus

View all literature mentions

MDA-MB-231 (cell line)

RRID:CVCL_0062

Cell line MDA-MB-231 is a Cancer cell line with a species of origin Homo sapiens (Human)

View all literature mentions

DLD-1 (cell line)

RRID:CVCL_0248

Cell line DLD-1 is a Cancer cell line with a species of origin Homo sapiens (Human)

View all literature mentions