KYSE-150 CRISPR cell line | 100% guarantee | High-efficiency

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KYSE-150 CRISPR cell line | 100% guarantee | High-efficiency

KYSE-150 cells are poorly differentiated esophageal adenocarcinoma cells that were isolated from the neck esophagus of a 49-year-old Japanese female patient who had received radiotherapy. When researchers discovered this cell line, the patient's cancer tissue had invaded the adjacent tissues. The KYSE-150 cell line is an epithelial cell with an adherent monolayer. It has been reported that cells carry increased oncogenes c-erb-B (8 times) and cyclin D1 (4 times) and that cells can form tumors in nude mice, so KYSE-150 cell line is often used in cancer research involving gene editings, such as CRISPR gene knockout/knock-in and point mutation.

 Applications:

1. EZH2 knockout in KYSE-150 cells altered PSMA3-AS1-induced proliferation and migration in esophageal cancer cells

PSMA3-AS1 expression is up-regulated and positively correlated with tumor size and metastasis in esophageal cancer patients. To further explore the biological roles of PSMA3-AS1 in esophageal cancer cells, researchers established stable CRISPR PSMA3-AS1-overexpressing cell lines via lentiviral infection in KYSE150 and KYSE450 cell lines (which have low PSMA3-AS1 expression) and validated the up-regulation of PSMA3-AS1 by RT-qPCR.

CRISPR/Cas9 gene-editing of EZH2 was successfully performed in KYSE150 and KYSE450cells as confirmed by a significant reduction in EZH2 protein expression. CCK-8 and colony formation assays showed that PSMA3-AS1 overexpression did not affect the proliferation of esophageal cancer cells with CRISPR EZH2 knocked out (Figure 6B and 6C). According to wound healing and transwell migration assays, wounding healing and cell migration were not increased in ESCC EZH2-knock out KYSE-150 cells compared to negative control cells.

Clinical-pathological characteristics illustrated that increased expression of PSMA3-AS1 was positively associated with distant metastasis, larger tumor sizes, and a worse prognosis for ESCC patients.


 2. CRISPR Over-expressed KYSE-150 cells increase chemo-resistance in ESCC

Whole-genome clustered regularly interspaced short palindromic repeats/CRISPR associated (CRISPR/Cas)-based a lentiviral library is a powerful tool for genome-scale gain-of-function or loss-of-function screening. This system has been proved to be highly effective in identifying drug-resistant genes in vitro. Shalem, Kurata, and Joung have screened out essential genes for drug resistance in melanoma and AML using the CRISPR knockout library. Some researchers attempted to combine the CRISPR library screening strategy with RNA sequencing technology in KYSE-150 Cells to explore the critical genes and potential mechanism for chemo-resistance in ESCC.

To increase the chance of identifying essential genes involved in PTX resistance, an integrated analysis was performed to combine EN-genes in genome-scale CRISPR screening and differentially expressed genes (DE-genes) in KYSE-150 cell line.

Researchers evaluated the potential of CDKN1A, ELAVL2, and TSPAN4 to promote chemo-resistance in ESCC cells KYSE-150. The results showed that overexpression of CDKN1A, ELAVL2, or TSPAN4 could significantly increase the resistance to PTX in both KYSE-180 and KYSE-150 cells. Moreover, overexpressed CDKN1A, ELAVL2, or TSPAN4 could also contribute to DDP-resistance in KYSE-150 cells.


 3. CRISPR/Cas9 mediated knockout of DEPTOR in KYSE-510 cells significantly promoted cellular proliferation, migration, and invasion

Researchers found that the expression of DEPTOR negatively regulates the tumorigenic activities of ESCC cell lines. Furthermore, ectopic DEPTOR expression caused significant suppression of the cellular proliferation, migration, and invasion of KYSE150 cells, which has the lowest expression level of DEPTOR. Meanwhile, CRISPR/Cas9 mediated knockout of DEPTOR in KYSE-510 cells significantly promoted cellular proliferation, migration, and invasion. Besides, in vivo assays further revealed that tumor growth was significantly inhibited in xenografts with ectopic DEPTOR expression as compared to untreated KYSE150 cells, and was markedly enhanced in DEPTOR knockout KYSE-510 cells.

In this study, scientists generated stable cell lines that either overexpressing DEPTOR or genetic ablation of endogenous expression of DEPTOR. Since KYSE-150 expresses the lowest endogenous level of DEPTOR among the three cell lines, they stably overexpressed DEPTOR in KYSE-150 cells (pcDNA3.1-DEPTOR). For the same consideration, KYSE-510 cells that express the highest level of DEPTOR were treated with CRISPR/Cas9 system to knockout of DEPTOR (CRISPR-DEPTOR). After the generation of cell lines, pcDNA3.1-DEPTOR displayed a reduced cell proliferation rate as compared to that of KYSE-150 parental cells and empty vector-transfected cells, while CRISPR-DEPTOR cells proliferated significantly faster than control KYSE-510 cells. Furthermore, pcDNA3.1-DEPTOR cells also showed reduced migration. Thus, these results suggested DEPTOR indeed regulates cell proliferation, migration, and invasion in ESCC cells.

Ubigene Biosciences is co-founded by biological academics and elites from China, the United States, and France. We are located in Guangzhou Science City, which serves as a global center for high technology and innovation. Ubigene Biosciences has 1000㎡ office areas and laboratories, involving genome editing, cell biology technology, and zebrafish research. We provide products and services for plasmids, viruses, cells, and zebrafish. We aim to provide customers with better gene-editing tools for cell or animal research.

We developed CRISPR-U™ and CRISPR-B™(based on CRISPR/Cas9 technology) which is more efficient than general CRISPR/Cas9 in double-strand breaking, CRISPR-U™ and CRISPR-B™ can greatly improve the efficiency of homologous recombination, easily achieve knockout (KO), point mutation (PM) and knockin (KI) in vitro and in vivo. 


According to customer needs, Yuanjing Biotechnology designs a stable gene transfer knockout program based on the target gene.

Scheme 1: Small-segment gene knockout scheme, gRNA is set in the introns at both ends of exon 2, and the number of coding bases of the knockout exon is not 3 times, and the knockout can cause frame shift.

Scheme 2: Frameshift gene knockout scheme, gRNA is set on the exon, the number of missing bases is not 3 times, and frameshift mutation can occur after knockout.
Scheme 3: Large-segment gene knockout scheme, knock out the coding sequence of the entire gene to achieve the effect of large-segment knockout.

Ubigene Biosciences is co-founded by biological academics and elites from China, the United States, and France. We are located in Guangzhou Science City, which serves as a global center for high technology and innovation. Ubigene Biosciences has 1000㎡ office areas and laboratories, involving genome editing, cell biology technology, and zebrafish research. We provide products and services for plasmids, viruses, cells, and zebrafish. We aim to provide customers with better gene-editing tools for cell or animal research.

We developed CRISPR-U™ and CRISPR-B™(based on CRISPR/Cas9 technology) which is more efficient than general CRISPR/Cas9 in double-strand breaking, CRISPR-U™ and CRISPR-B™ can greatly improve the efficiency of homologous recombination, easily achieve knockout (KO), point mutation (PM) and knockin (KI) in vitro and in vivo. 


References:

1. Nødvig CS, Hoof JB, Kogle ME, et al. Efficient oligonucleotide mediated CRISPR-Cas9 gene editing in Aspergilli. Fungal Genet Biol. 2018;115:78-89

2.  Nødvig CS, Nielsen JB, Kogle ME, Mortensen, UH. A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. PLoS One. 2015;10(7):e0133085. Published 2015 Jul 15.

3. Vanegas, K.G., Jarczynska, Z.D., Strucko, T. et al. Cpf1 enables fast and efficient genome editing in Aspergilli. Fungal Biol Biotechnol 6, 6 (2019).

4. Nayak T, Szewczyk E, Oakley CE, et al. A versatile and efficient gene-targeting system for Aspergillus nidulans. Genetics. 2006;172(3):1557-1566. 


Genome Editing Platform

——Focusing on the Application of CRISPR-U™ and CRISPR-B™ Gene Editing Technology
1. Provides various types of gene-editing vectors for different species.
2. Provides different virus packaging services, including lentiviruses, adenoviruses and adeno-associated viruses.3. Provides high-quality services for gene knockout, point mutation and knockin cell lines

Cell Biology Platform
——Focusing on primary cell
1. Provides over 400 types of primary cells.

2. Provides culture strategies and related products for different cell types.3. Provides cell biology-related services such as cell isolation, extraction and validation.