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CRISPR-U™ Gene Knockout Cell Line Strategy

FH 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 FH Knockout model in cell line by CRISPR-U™-mediated genome engineering.
Target gene info
Official symbol
FH
Gene id
2271
Organism
Homo sapiens
Gene type
protein-coding
Official full symbol
fumarate hydratase
Also known as
FMRD, HLRCC, HsFH, LRCC, MCL, MCUL1
Genomic regions
Chromosome 1
Summary
The protein encoded by this gene is an enzymatic component of the tricarboxylic acid (TCA) cycle, or Krebs cycle, and catalyzes the formation of L-malate from fumarate. It exists in both a cytosolic form and an N-terminal extended form, differing only in the translation start site used. The N-terminal extended form is targeted to the mitochondrion, where the removal of the extension generates the same form as in the cytoplasm. It is similar to some thermostable class II fumarases and functions as a homotetramer. Mutations in this gene can cause fumarase deficiency and lead to progressive encephalopathy.
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 FH gene is evaluated as high lethality riskin cell line, and the CRISPR gene-editing strategy design is Unknown.
Knockout project comprehensive difficulty is thus assessed as Unknown. KO experiment is not recommended.
Red Cotton™ Notes Gene FH had been KO in hek293t cell line.
EZ-editor™ Gene Dependency
EZ-editor™ Gene Expression Level
EZ-editor™ Gene Copy Number

EZ-editor™ Gene Dependency

Result

The FH gene in cell line is evaluated as

Gene lethality score

Dependent Cell Lines Gene Dependency 0 0.5 1

Lethality score of FH in different cells

13.7%>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 0.1% cells with low expression level, 99.9% cells with medium expression level, 0.1% cells with high expression level of FH 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 FH in different cells

0.1%99.9%0.1%
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 47.8% cells with low copy number, 46.2% cells with medium copy number, 6.0% cells with high copy number of FH 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 50 100 150 200 250 300 350 400 450Cell Lines

Copy number of FH in different cells

47.8%46.2%6.0%
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。