CRISPR library screening is a high-throughput gene screening protocol based on CRISPR Cas9 technology, which infects target cells with low MOI by constructing a library containing thousands of sgRNAs and cloning these sgRNAs into lentiviral vectors. This approach ensures that each cell is infected with only one type of sgRNA, thus enabling functional screening of genes corresponding to different sgRNAs. We recommend the three best-selling CRISPR libraries in stock this month, and attach the interpretation of the relevant research literature of the library, hoping to contribute to everyone's scientific research path
OneMetabolismGenesLibraryDecodingGenes associated with triple-negative breast cancer metastasis
Original link: Breast cancer is the most common malignant tumor that endangers women's health worldwide, and its high incidence and mortality have attracted increasing attention, among which triple-negative breast cancer accounts for 12-17% of all breast cancers. Metabolic reprogramming plays an important role in tumor development, and multiple genes involved in lipid metabolism may also be involved in tumor development, but the role of most lipid molecules and enzymes in the molecular mechanism of triple negative metastasis in breast cancer remains unclear.
To study the association of lipid metabolism-related genes with breast cancer, the researchers used:Lipid metabolism gene CRISPR knockout librariesScreening was carried out to identify glycerol diacylkinase (DGKZ) as a potential premetastatic gene, and the knockout of DGKZ in triple-negative breast cancer cell lines showed that it could significantly inhibit metastatic behavior in vitro and in vivo, while overexpression of DGKZ would increase the metastatic potential of cell lines, confirming that high expression of DGKZ was associated with tumor progression and poor prognosis in patients. The lipid metabolism library has successfully screened genes related to triple-negative breast cancer metastasis, which plays a key role in exploring tumorigenesis and progression, and provides potential targets for the development of new strategies.
Fig.1 Lipid metabolism library screened the gene DGKZ related to triple negative breast cancer metastasis
2. PeopleTranscription factorsLibrary screeningEnteroendocrine cellsDominant inhibitors
Enteroendocrine cells (EECs) are hormone-producing cells that grow within the epithelium of the stomach, small intestine, and colon, and regulate various aspects of metabolic activity, including insulin levels, satiety, gastrointestinal secretion, and motility. However, the differentiation process of intestinal endocrine cells is not fully understood.
To study the differentiation process of intestinal endocrine cells, researchers used:Human transcription factorscrisprKnock out the libraryAll transcription factors in small intestinal organoids were screened, and it was found that ZNF800 is a dominant inhibitor controlling the fate of endocrine cells, which restricts the differentiation of enterochromaffin cells by directly regulating endocrine transcription factor networks such as PAX4. The discovery of the dominant inhibitor of endocrine cells, ZNF800, by CRISPR gene knockout library technology, not only increases our understanding of intestinal endocrine cell differentiation, but also provides new potential targets for the future.
Fig.2 Human transcription factor knockout library screened the dominant inhibitor ZNF800 in enteroendocrine cells
IIIrnacombinedEgg whitesLibrary revealedRegulationpd-l1Key factors in tumor immune evasion
Original link: RNA binding proteins (RBPs) are a class of proteins that can bind to RNA, which can recognize and bind specific RNA sequences, thereby regulating gene expression, transcription, translation and other processes. RBP dysfunction has been observed in various cancers in previous studies, however, it is unclear whether specific RBPs are involved in tumor immune evasion by regulating programmed deathligand-1 (PD-L1).
To do this, the researchers conducted:rna-binding proteinKnockoutLibrary screeningIt was found that density-regulated restart and release factor (DENR) is a protein that can regulate PD-L1, and further studies have found that the loss of DENR will lead to a decrease in the expression of PD-L1, thereby affecting tumor growth and immune cell response. DENR regulates the expression of PD-L1 by antagonizing the translational inhibition of three successive upstream open reading frames (UORFS) and promoting the translation of Janus kinase 2 (JAK2) and the activation of the IFN-JAK-STAT signaling pathway. The systematic discovery and identification of new PD-L1 regulators DENR using RBP CRISPR Cas9 library screening provides a new perspective for understanding the mechanism of tumor immune evasion, and also provides a new idea for the development of new tumor methods.
Fig.3 CRISPR Cas9 library screened DENR, a key regulator of RBP
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