Rad51 Gene Editing

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Rad51 Gene Editing    

RAD51, DNA repair protein RAD51 homolog 1, its family members are highly similar to bacterial RecA and Saccharomyces cerevisiae Rad51, and are known to be involved in the homologous recombination and repair of DNA. The complete and accurate replication of the genome is essential to maintain genome integrity. As the central homologous recombination (HR) protein, this protein can form helical nucleoprotein filaments on tracts of single strand DNA, and interact with the ssDNA-binding protein RPA and RAD52, and it is thought to play central roles in homologous pairing, recombinational repair and strand transfer of DNA in eukaryotes. One additional thing to note is that Rad51 plays an important part in homologous strand exchange, a key step in DNA repair through HR. Rad51 is also involved in interstrand cross-link repair. This encoded protein binds to single and double-stranded DNA and exhibits DNA-dependent ATPase activity. Rad51 catalyzes the recognition of homology and strand exchange between homologous DNA partners to form a joint molecule between a processed DNA break and the repair template. It also binds to single-stranded DNA in an ATP-dependent manner to form nucleoprotein filaments which are essential for the homology search and strand exchange. Replication forks face many obstacles that can result in replication fork stalling or replication fork collapse. Besides its role in DNA double strand breaks (DSBs) repairing, a non-enzymatic function of RAD51 promotes replication fork regression, a process that has also been referred to as replication fork reversal, which involves branch migration in the direction opposite to replication to form a Holliday junction-containing chicken foot structure. In the same study, Jennifer M. Mason, etc. shows that RAD51 promotes replication restart by both strand exchange-dependent and strand exchange-independent mechanisms, and protects newly synthesized DNA from degradation.

RAD51 interacts (directly or indirectly) with a large number of proteins involved in DNA repair and the cell cycle. This RAD51 protein is found to interact with p53, BRCA1 and BRCA2, which may be important for the cellular response to DNA damage. Cooperation between the DNA recombinase RAD51 and the breast cancer susceptibility protein BRCA2 is crucial for the repair of double-strand DNA breaks. BRCA2 is shown to regulate both the intracellular localization and DNA-binding ability of this protein. Part of a PALB2-scaffolded HR complex containing BRCA2 and RAD51C, is thought to play a role in DNA repair by HR and regulating the copy number of mitochondrial DNA under conditions of oxidative stress in the presence of RAD51C and XRCC3. Loss of these controls following BRCA2 inactivation may be a key event leading to genomic instability and tumorigenesis. Multiple transcript variants encoding different isoforms have been found for this gene.

Rad51 Gene EditingFig. 1 RAD51, BRCA2 and DNA repair: a partial resolution (Christopher J Lord & Alan Ashworth, 2007)

It is well known that deficiencies in DNA repair can lead to carcinogenesis. Loss of RAD51 function would lead to accumulation of DNA damage and, therefore, to increased cancer risk. There is evidence to support a key role for RAD51 in breast cancer and breast tumourigenesis. Researchers did not detect any variation in the coding region of RAD51, which may further imply the importance of maintaining the structural integrity of such a vital component of the DNA repair pathway.

CancerOver or Under expressionEvaluation method
Breast cancer (invasive ductal)Over-expressionImmunohistochemistry
Breast cancer (BRCA1 deficient)Over-expressionmessenger RNA
Breast cancer (progesteron receptor negative)Over-expressionmessenger RNA
Breast cancerUnder-expressionImmunohistochemistry
Pancreatic cancerOver-expressionImmunohistochemistry
Pancreatic cancerOver-expressionImmunohistochemistry
Head and neck squamous cancersOver-expressionImmunohistochemistry
Prostate cancerOver-expressionImmunohistochemistry
Non-small-cell lung cancerOver-expressionImmunohistochemistry
Soft tissue sarcomaOver-expressionImmunohistochemistry
Esophageal squamous cell cancerOver-expressionImmunohistochemistry
Renal cell carcinomaUnder-expressionWestern (protein) blotting and mRNA

Table 1. RAD51 expression in sporadic cancers

Genome Editing is revolutionizing biomedical research, due to its high efficiency, ease-of-use, and relatively low cost. CRISPR/Cas9 PlatformCB, a global biotechnological company specializing in gene editing, is dedicated to offering comprehensive CRISPR/Cas9 gene editing products and services for academic research, biotech research and pharmaceutical drug discovery. With products for CRISPR-Cas9 clones, validation and screening kits, pre-made transfected stable cell lines, and more for precision genome editing, our complete solutions for genome editing provide you with a wealth of tools to help you every step of the way in your genome editing workflow.

  • Gene Editing Cell Line Generation

Our specialists will provide with helpful tips on designing and using gRNA and Cas9 constructs, CRISPR libraries, designing KO mammalian cell lines, and more!

ServiceDetailsCell Line Options *
Knockout Cell Line ServiceSingle gene knockout cell lineAvailable for any transfection-suitable cell lines, including A549, CHO-K1, HEK293, HEK293T, HCT116, MCF7, MDCK, U937, RPMI 8866, HT-29, MDA-MB-231, 4T1, A20, etc.
Transfection-based
Customer specifies gene/locus region and cell line of interest*
Customized Knockout Cell Line ServiceSingle or multiplex genes editing knockout cell lineAny cancer cell line
Transfection/Lentivirus/Ribonucleoprotein-based
Customer specifies gene/locus region and cell line of interest*

* It is preferred that customers provide their own cell lines, despite available cell lines can be purchased by Creative Biogene for an additional fee.

Related Details

Guide RNA Design:

Our expert scientists from Creative Biogene will optimize your guide RNA sequence for maximized efficiency and minimal off-target effects using best design algorithms.

Materials Needed from Clients:

Target gene name.

Target gene sequence, if the whole genome sequence is unavailable.

Quality Control

Sequence Chromatogram

Quality Assurance Certificate

Quantitative RT-PCR Validation (Optional)

Western Blot Validation (Optional)

Deliverables

Knockout cell lines validated by sequencing

Delivery Time

Starting from 4 weeks

  • Gene Editing Animal Model Generation
ServiceDetailsDelivery TimeOptions *
Gene Knockout Mouse GenerationConditional Knockout Mouse
Conventional Knockout Mouse
Point Mutation Mouse
Starting from 6 weeks 
Gene Knockin Mouse GenerationCRISPR/Cas9 Knockin Mouse
Rosa26 Knockin Mouse
Starting from 6 weeksAny other target gene knockin mouse generation
Custom Animal ModelsCustom Rat Model
Custom Rabbit Model
Custom Zebrafish Modell
Custom C. elegans Model
Custom Drosophila Model
Starting from 6 weeksAny other animal models according to our client's requirements

CRISPR/Cas9 PlatformCB is devoted to providing the best gene editing services to accelerate the achievement of your research goals. We are committed to providing the most reliable and effective research services and products with excellent quality management and quality assurance capacity. If you have any questions, please feel free to contact us.

Related Products at CRISPR/Cas9 PlatformCB

CATALOG NO.PRODUCT NAMEPRODUCT TYPEINQUIRY
CSC-RT1456Human RAD52 Knockout Cell Line-HeLaKnockout Cell LineInquiry

Legal Statement of CRISPR/Cas9 PlatformCB, Services and Products

The products and the reagents generated from these services shall be only used as tools for research purposes, and shall exclude (a) any human or clinical use, including, without limitation, any administration into humans or any diagnostic or prognostic use, (b) any human germline modification, including modifying the DNA of human embryos or human reproductive cells, (c) any in vivo veterinary or livestock use, or (d) the manufacture, distribution, importation, exportation, transportation, sale, offer for sale, marketing, promotion or other exploitation or use of, or as, a testing service, therapeutic or diagnostic for humans or animals.

References

  1. Kolinjivadi, A. M. et al. Moonlighting at replication forks—a new life for homologous recombination proteins BRCA1, BRCA2 and RAD51. FEBS Lett. 47, 222 (2017).
  2. Berti, M. & Vindigni, A. Replication stress: getting back on track. Nat. Struct. Mol. Biol. 23, 103-109 (2016).
  3. Zellweger, R. et al. Rad51-mediated replication fork reversal is a global response to genotoxic treatments in human cells. J. Cell Biol.208, 563-579 (2015).
  4. Mijic, S. et al. Replication fork reversal triggers fork degradation in BRCA2-defective cells. Nat. Commun. 8, 859 (2017).
  5. Thangavel, S. et al. DNA2 drives processing and restart of reversed replication forks in human cells. J. Cell Biol. 208, 545-562 (2015).
For research use only. Not intended for any clinical use.

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