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CRISPR-U™ Gene Knockout Cell Line Strategy

JAK2 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 JAK2 Knockout model in cell line by CRISPR-U™-mediated genome engineering.
Target gene info
Official symbol
JAK2
Gene id
3717
Organism
Homo sapiens
Gene type
protein-coding
Official full symbol
Janus kinase 2
Also known as
JTK10
Genomic regions
Chromosome 9
Summary
This gene encodes a non-receptor tyrosine kinase that plays a central role in cytokine and growth factor signalling. The primary isoform of this protein has an N-terminal FERM domain that is required for erythropoietin receptor association, an SH2 domain that binds STAT transcription factors, a pseudokinase domain and a C-terminal tyrosine kinase domain. Cytokine binding induces autophosphorylation and activation of this kinase. This kinase then recruits and phosphorylates signal transducer and activator of transcription (STAT) proteins. Growth factors like TGF-beta 1 also induce phosphorylation and activation of this kinase and translocation of downstream STAT proteins to the nucleus where they influence gene transcription. Mutations in this gene are associated with numerous inflammatory diseases and malignancies. This gene is a downstream target of the pleiotropic cytokine IL6 that is produced by B cells, T cells, dendritic cells and macrophages to produce an immune response or inflammation. Disregulation of the IL6/JAK2/STAT3 signalling pathways produces increased cellular proliferation and myeloproliferative neoplasms of hematopoietic stem cells. A nonsynonymous mutation in the pseudokinase domain of this gene disrupts the domains inhibitory effect and results in constitutive tyrosine phosphorylation activity and hypersensitivity to cytokine signalling. This gene and the IL6/JAK2/STAT3 signalling pathway is a therapeutic target for the treatment of excessive inflammatory responses to viral infections. Alternative splicing results in multiple transcript variants encoding distinct 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 JAK2 had been KO in hela cell line.
EZ-editor™ Gene Dependency
EZ-editor™ Gene Expression Level
EZ-editor™ Gene Copy Number

EZ-editor™ Gene Dependency

Result

The JAK2 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 JAK2 in different cells

0.5%>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 3.5% cells with expression level below cutoff, 94.6% cells with low expression level, 1.9% cells with medium expression level of JAK2 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 JAK2 in different cells

3.5%94.6%1.9%
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 74.5% cells with low copy number, 21.5% cells with medium copy number, 4.0% cells with high copy number of JAK2 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 JAK2 in different cells

74.5%21.5%4.0%
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 5000+ 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。