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CRISPR-U™ Gene Knockout Cell Line Strategy

PTEN Gene Knockout Strategy

CRISPR-U™ technology (CRISPR based), developed by Ubigene, is more efficient than general CRISPR/Cas9 technology in double-strand breaking and homologous recombination. With CRISPR-U™, Ubigene has successfully edited over 3000 genes on more than 200 types of cell lines.
Objective
To create a Human PTEN Knockout model in cell line by CRISPR-U™-mediated genome engineering.
Target gene info
Official symbol
PTEN
Gene id
5728
Organism
Homo sapiens
Gene type
protein-coding
Official full symbol
phosphatase and tensin homolog
Also known as
10q23del, BZS, CWS1, DEC, GLM2, MHAM, MMAC1, PTEN1, PTENbeta, TEP1
Genomic regions
Chromosome 10
Summary
This gene was identified as a tumor suppressor that is mutated in a large number of cancers at high frequency. The protein 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. It negatively regulates intracellular levels of phosphatidylinositol-3,4,5-trisphosphate in cells and functions as a tumor suppressor by negatively regulating AKT/PKB signaling pathway. 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.
Strategy Summary
This gene has 0 protein coding transcripts:
Frame-shift
Fragment A
Fragment B
Strategy

Project Comprehensive Difficulty Assessment

According to the Red Cotton database: the CRISPR gene-editing strategy design is Unknown.
Knockout project comprehensive difficulty is thus assessed as Unknown.
Red Cotton™ Notes Gene PTEN had been KO in hct116 cell line.
EZ-editor™ Gene Dependency
EZ-editor™ Gene Expression Level
EZ-editor™ Gene Copy Number

EZ-editor™ Gene Dependency

Result

The PTEN gene you inquire is evaluated as high risk in 1% cell line. Cell line is not selected, unable to assess the accurate risk level, for reference only.

Gene lethality score

Dependent Cell Lines Gene Dependency 0 0.5 1

Lethality score of PTEN in different cells

1.1%>0.5
All cells
0
Non-essential gene
0.5
Boundary score
1
Essential gene

EZ-editor™ Gene Expression Level

Result

In all cell lines, there is 1.3% cells with expression level below cutoff, 7.9% cells with low expression level, 90.8% cells with medium expression level of PTEN gene. Cell line is not selected, unable to assess the accurate expression level, for reference only.

Gene Expression

Cell Lines Frequency log2(TPM+1) 0 2 4 6 8 10 12

TPM of PTEN in different cells

1.3%7.9%90.8%
All cells
0~0.5
Below cutoff
0.5~10
Low
10~1000
Medium
>1000
High

EZ-editor™ Gene Copy Number

Result

In all cell lines, there is 70.8% cells with low copy number, 27.2% cells with medium copy number, 2.0% cells with high copy number of PTEN gene. Cell line is not selected, unable to assess the accurate copy number, for reference only.

Gene Copy Number

Copy number 0 1 2 3 4 5 6 7 0 100 200 300 400 500 600Cell Lines

Copy number of PTEN in different cells

70.8%27.2%2.0%
All cells
0~2
Low
2~4
Medium
>4
High
Work flow
Ubigene Red Cotton Workflow

Ubigene - Make genome editing easier

Ubigene is an international high-technology enterprise focused on gene-editing cells. Our exclusive CRISPR-U™ technology has 10-20 times more efficient editing than traditional methods, easily achieving gene knockout, point mutation, and knock-in. Based on CRISPR-U™ technology, Ubigene has accumulated over 6000 successful gene-editing cases from more than 300 cell lines including iPSC and ESC, and has established a KO Cell Line Bank with 4500+ KO cell lines and Red Cotton™ gRNA Plasmid Bank with 10000+ gRNA plasmids available in stock.

Ubigene focuses on technological innovation and product development, of which EZ-editor™ series products that cover the whole workflow of gene-editing keep improving. Ubigene will move on toward our goal of "Make genome editing easier" and we won't stop!

Cell products

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红棉系统新技能上线啦!基因风险评估:“火眼金睛”辨基因,致死风险一秒知。评估结果可在Red Cotton Assessment模块查看哦。