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PTEN (phosphatase and tensin homolog), also referred to as MMAC1, a protein encoded by the gene in humans, is a multi-functional tumor suppressor that is very commonly lost in human cancer, as well as the mutations of this gene are a step in the development of those diseases. PTEN has diverse functions in mammalian cells. This gene was identified as a tumor suppressor that is mutated in a large number of cancers at high frequency, through the action of its phosphatase protein product. This phosphatase is involved in the regulation of the cell cycle, preventing cells from growing and dividing too rapidly, and widely expressed throughout the body.
Figure 1. Overview of the PTEN network (M Keniry et al., 2008)
The protein product encoded by this gene is a phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase. It contains a tensin like domain as well as a catalytic domain similar to that of the dual specificity protein tyrosine phosphatases. Unlike most of the protein tyrosine phosphatases, this protein preferentially dephosphorylates phosphoinositide substrates. PTEN inhibits growth factor signals sent through PI3 kinase. It negatively regulates intracellular levels of phosphatidylinositol-3,4,5-trisphosphate(PIP3) in cells and functions as a tumor suppressor, negatively regulating AKT/PKB signaling pathway by dephosphorylating the PIP3, resulting in the biphosphate product PIP2. Loss of PTEN expression results in the downstream activation (phosphorylation) of AKT, an inhibitor of apoptosis and promoter of cell proliferation. The use of a non-canonical (CUG) upstream initiation site produces a longer isoform that initiates translation with a leucine, and is thought to be preferentially associated with the mitochondrial inner membrane. This longer isoform may help regulate energy metabolism in the mitochondria. A pseudogene of this gene is found on chromosome 9. Alternative splicing and the use of multiple translation start codons results in multiple transcript variants encoding different isoforms.
Together with the discovery of the PTEN gene in a loss of heterozygosity (LOH) -prone region of chromosome 10, mutational analysis immediately revealed frequent PTEN point mutations in commonly used cancer cell lines, and primary tumor samples. PTEN is very commonly lost in human cancer, observed in prostate cancer, glioblastoma, endometrial, lung and breast cancer to varying degrees, which at times exhibit homozygous deletion of PTEN. However, the majority of primary prostate cancer cases with genetic alterations in PTEN harbor loss of heterozygosity at the PTEN locus without mutations in the remaining allele. During tumor development, mutations and deletions of PTEN occur that inactivate its enzymatic activity leading to increased cell proliferation and reduced cell death. Most often complete inactivation of this gene is associated with late-stage, aggressive, usually metastatic tumors. A complete loss of PTEN function in early-stage tumors appears to be unique for ovarian and endometrial cancers. Moreover, diseases associated with PTEN include Cowden Syndrome 1, and Macrocephaly/Autism Syndrome. As all mentioned above, it is a part of the PI3K/AKT/mTOR pathway and mTOR inhibitors have been relatively ineffective in treating patients with PTEN loss. PTEN has attracted great interest from various research fields because of its diverse physiological functions. The functional diversity of PTEN demands a collection of delicate regulatory mechanisms, including transcriptional and posttranslational regulation in a tissue- and context-dependent manner. New appoaches using microRNAs are currently being investigated. PTEN is one of the targets for drug candidates such as the oncomiR, MIRN21.
PTEN Gene Editing Service
CRISPR/Cas9 PlatformCB, a global leading biotechnological company specializing in gene editing, is dedicated to offering comprehensive CRISPR/Cas9 gene editing services and products for academic research, biotech research and pharmaceutical drug discovery. With deep gene editing knowledge and extensive experience in experimental operation and data processing, we help you effectively control target genes knockout/knockin/point mutation in cells or animals via CRISPR/Cas9 technology.
Our professional scientists have successfully implemented PTEN CRISPR/Cas9 gene edited in both easy-to-transfect cell lines and hard-to-transfect cells. To support your projects, we will offer you full-length custom PTEN gene editing service form strategy design to final stable cells. Our PTEN gene editing cell line generation services include:
➢ gRNA design and synthesis
➢ Transfect the cell lines you’re interested
➢ Select the high expression cells and sort monoclonal cell
➢ Validate the knockout/knockin/point mutation of PTEN by PCR and sequencing
➢ Provide cryogenic preserved vials of stable cells and final reports
Typically, we develop CRISPR-mediated gene editing cell lines including HEK239T, Hela, HepG2, U87, but we can use other cell lines according to your requirements.
Other host cell lines available: Ba/F3, CHO, MDA-MB-453, MDA-MB-231NIH3T3, T47D, Neuro2a, MCF7, RKO, K562, RAW264.7, etc.
CRISPR/Cas9 PlatformCB also has extensive experience in incorporating CRISPR-Cas9 technology into animal models, which have been fully recognized by our clients. Tell us your projects’ needs, we can provide a one-stop-shop PTEN CRISPR/Cas9 gene editing animal service and guarantee at least 2 founders or 3 F1 animals with shorter turnaround time and lower price. Our PTEN gene editing animal model generation services include:
➢ PTEN gene conventional knockout animals
➢ PTEN gene conditional knockout animals
➢ PTEN point mutation animals
➢ PTEN knockin animals
Alternative species: mouse, rat, rabbit, zebrafish, C. elegans, etc.
CRISPR/Cas9 PlatformCB is devoted to providing the best gene editing services to accelerate the achievement of your research goals. We are committed to providing the most reliable and effective research services and products with excellent quality management and quality assurance capacity. If you have any questions, please feel free to contact us.
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Legal Statement of CRISPR/Cas9 PlatformCB, Services and Products：
The products and the reagents generated from these services shall be only used as tools for research purposes, and shall exclude (a) any human or clinical use, including, without limitation, any administration into humans or any diagnostic or prognostic use, (b) any human germline modification, including modifying the DNA of human embryos or human reproductive cells, (c) any in vivo veterinary or livestock use, or (d) the manufacture, distribution, importation, exportation, transportation, sale, offer for sale, marketing, promotion or other exploitation or use of, or as, a testing service, therapeutic or diagnostic for humans or animals.