CRISPR/Cas9 mediated KYSE-150 cell line | Knockout, Knockin & Point mutation

 
GUANGZHOU, China - Dec. 1, 2020 - PRLog -- 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

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

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


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.


See the full article at https://ubigene.us/about/article/2886.html

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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.

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