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CRISPR-U™ Gene Knockout Cell Line Strategy

BRCA1 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 BRCA1 Knockout model in cell line by CRISPR-U™-mediated genome engineering.
Target gene info
Official symbol
BRCA1
Gene id
672
Organism
Homo sapiens
Gene type
protein-coding
Official full symbol
BRCA1 DNA repair associated
Also known as
BRCAI, BRCC1, BROVCA1, FANCS, IRIS, PNCA4, PPP1R53, PSCP, RNF53
Genomic regions
Chromosome 17
Summary
This gene encodes a 190 kD nuclear phosphoprotein that plays a role in maintaining genomic stability, and it also acts as a tumor suppressor. The BRCA1 gene contains 22 exons spanning about 110 kb of DNA. The encoded protein combines with other tumor suppressors, DNA damage sensors, and signal transducers to form a large multi-subunit protein complex known as the BRCA1-associated genome surveillance complex (BASC). This gene product associates with RNA polymerase II, and through the C-terminal domain, also interacts with histone deacetylase complexes. This protein thus plays a role in transcription, DNA repair of double-stranded breaks, and recombination. Mutations in this gene are responsible for approximately 40% of inherited breast cancers and more than 80% of inherited breast and ovarian cancers. Alternative splicing plays a role in modulating the subcellular localization and physiological function of this gene. Many alternatively spliced transcript variants, some of which are disease-associated mutations, have been described for this gene, but the full-length natures of only some of these variants has been described. A related pseudogene, which is also located on chromosome 17, has been identified.
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 BRCA1 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.
EZ-editor™ Gene Dependency
EZ-editor™ Gene Expression Level
EZ-editor™ Gene Copy Number

EZ-editor™ Gene Dependency

Result

The BRCA1 gene in cell line is evaluated as

Gene lethality score

Dependent Cell Lines Gene Dependency 0 0.5 1

Lethality score of BRCA1 in different cells

32.6%>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 expression level below cutoff, 16.4% cells with low expression level, 83.5% cells with medium expression level of BRCA1 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 BRCA1 in different cells

0.1%16.4%83.5%
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 57.1% cells with low copy number, 37.7% cells with medium copy number, 5.2% cells with high copy number of BRCA1 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 BRCA1 in different cells

57.1%37.7%5.2%
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。