[FEB Bestselling] CRISPR Screen for Metabolic, Transcription Factor, RNA-Binding Protein
CRISPR screening is a high-throughput genetic screening by constructing a library containing many sgRNAs and cloning these sgRNAs into lentiviral vectors to infect target cells at low MOI. This approach ensures that each cell is infected by one sgRNA, thus enabling functional screening against genes corresponding to different sgRNAs. In this article, we recommend the three best-selling CRISPR libraries this month, and introduce corresponding literatures interpretation. Hope this could inspire your research!
Breast cancer is the most common malignant tumor endangering women's health worldwide, and its high morbidity and mortality have attracted increasing attention, with triple-negative breast cancer accounting for 12-17% of all breast cancers. Metabolic reprogramming plays an important role in tumor development, and a variety of genes involved in lipid metabolism may also be involved in tumor development, but the roles of most lipid molecules and enzymes in the molecular mechanisms of triple-negative metastasis in breast cancer remain unclear.
To study the association between lipid metabolism and breast cancer, researchers screened using Human CRISPR Metabolic Gene Knockout Library and identified glycerol diacyl kinase ζ (DGKZ) as a potential pre-metastatic gene. Knockout DGKZ in triple-negative breast cancer cell lines significantly inhibited metastatic behavior both in vitro and in vivo. Overexpression of DGKZ increased the metastatic potential of the cell lines. This shows that high DGKZ expression is associated with tumor progression and poor prognosis in patients. The Metabolic Gene Knockout library successfully screened genes associated with metastasis in triple-negative breast cancer, which plays a key role in exploring tumorigenesis and progression, and provides potential targets for the development of new therapeutic.
Fig.1 Results of Metabolic Gene Knockout Library screening. DGKZ, a metastasis-related gene in triple-negative breast cancer
Human Transcription Factor Knockout Library screening for enteroendocrine cell dominant repressors
Enteroendocrine cells (EECs) are hormone-producing cells that grow within the epithelium of the stomach, small intestine, and colon. EECs regulate various aspects of metabolic activity, including insulin levels, satiety, gastrointestinal secretion, and motility. However, the process of differentiation of enteroendocrine cells is not fully understood.
To study the differentiation process of intestinal endocrine cells, researchers screened all transcription factors in adult small intestinal organoids using Human Transcription Factor Knockout Library and found that ZNF800 is a dominant repressor controlling the fate of endocrine cells, which restricts Enterochromaffin cell differentiation through the direct regulation of a network of endocrine transcription factors such as PAX4. The discovery of the endocrine cell dominant repressor ZNF800 by CRISPR knockdown library technology not only increases our understanding of enteroendocrine cell differentiation, but also provides new potential targets for future therapies.
Fig 2. Human Transcription Factor Knockout Library screening for the enteroendocrine cell dominant suppressor ZNF800
RNA-binding protein library reveals key factor regulating immune escape
RNA-binding proteins (RBPs) are a class of proteins that are able to bind to RNA and recognize and bind specific RNA sequences, thereby regulating multiple processes such as gene expression, transcription, and translation. RBP dysfunction has been observed in various cancers in previous studies; however, whether specific RBPs are involved in tumor immune evasion through the regulation of programmed death ligand-1 (PD-L1) is unknown.
Researchers conducted an RNA-Binding Protein Pooled CRISPR Knockout Library screen and identified density-regulated re-initiation and release factor (DENR) as a protein capable of regulating PD-L1, and through further studies, found that knockout of DENR leads to reduced PD-L1 expression, which in turn affects tumor growth and immune cell response. DENR promotes the translation of Janus kinase 2 (Jak2) and IFNγ-JAK-STAT signaling pathways by antagonizing the translational repression of three consecutive upstream open reading frames (uORFs) and promotes translation of Janus kinase 2 (Jak2) and activation of the IFNγ-JAK-STAT signaling pathway, thereby regulating PD-L1 expression. The systematic discovery and identification of new PD-L1 regulator DENR using RBP CRISPR library screening provides new perspectives for understanding the mechanism of tumor immune escape and new ideas for developing new tumor therapeutic approaches.
Fig. 3 RBP CRISPR screen for key regulation factor DENR
EDITGENE offers aforementioned Metabolic, Transcription Factor, RNA-Binding Protein CRISPR libraries. Pre-made libraries deliver in 1 week, only $1600 get 100ug plasmid library. Explore our CRISPR libraries now>>
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