EZ-editor™ technology (CRISPR based), developed by Ubigene, is more efficient than general
CRISPR/Cas9 technology in double-strand breaking and homologous recombination. With EZ-editor™, Ubigene has
successfully edited over 10000 genes on more than 500 types of cell lines.
Objective
To create a Human BRCA2 Knockout
model in cell line by EZ-editor™-mediated genome engineering.
Inherited mutations in BRCA1 and this gene, BRCA2, confer increased lifetime risk of developing breast or ovarian cancer. Both BRCA1 and BRCA2 are involved in maintenance of genome stability, specifically the homologous recombination pathway for double-strand DNA repair. The largest exon in both genes is exon 11, which harbors the most important and frequent mutations in breast cancer patients. The BRCA2 gene was found on chromosome 13q12.3 in human. The BRCA2 protein contains several copies of a 70 aa motif called the BRC motif, and these motifs mediate binding to the RAD51 recombinase which functions in DNA repair. BRCA2 is considered a tumor suppressor gene, as tumors with BRCA2 mutations generally exhibit loss of heterozygosity (LOH) of the wild-type allele.
Strategy Summary
This gene has 6 protein coding transcripts：
Transcript
Length(nt)
Protein
Length(aa)
Protein name
Transcript type
Transcript name
ccds
NM_001432077
11863
NP_001419006
3418
breast cancer type 2 susceptibility protein
3
transcript variant 7
NM_000059
11954
NP_000050
3418
breast cancer type 2 susceptibility protein
3
transcript variant 1
CCDS9344.1
NM_001406720
11903
NP_001393649
3401
breast cancer type 2 susceptibility protein
3
transcript variant 2
NM_001406719
11858
NP_001393648
3386
breast cancer type 2 susceptibility protein
3
transcript variant 3
NM_001406721
7022
NP_001393650
1774
breast cancer type 2 susceptibility protein
3
transcript variant 4
NM_001406722
5800
NP_001393651
1279
breast cancer type 2 susceptibility protein
3
transcript variant 5
Frame-shift
Fragment A
Fragment B
gRNA Detail
Strategy
Project Comprehensive Difficulty Assessment
According to the Red Cotton database: the BRCA2 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
BRCA2
had been KO in pc-3m cell line.
In all cell lines, there is
1.8% cells with expression level below cutoff,
83.1% cells with low expression level,
15.1% cells with medium expression level
of BRCA2 gene.
Cell line is not selected, unable to assess the accurate expression level, for reference only.
In all cell lines, there is
72.2% cells with low copy number,
24.5% cells with medium copy number,
3.3% cells with high copy number
of BRCA2 gene.
Cell line is not selected, unable to assess the accurate copy number, for reference only.
Ubigene is an international high-technology enterprise focused on gene-editing cells. Our exclusive EZ-editor™ technology has 10-20 times more efficient editing than traditional methods, easily achieving gene knockout, point mutation, and knock-in. Based on EZ-editor™ 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!
EZ-editor™ Gene Dependency
EZ-editor™ Gene Expression Level 
EZ-editor™ Gene Copy Number 
