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Custom Gene Targeting Vector Synthesis


Item Cat No.: BCCRGT

Application: Genome editing

Reactivity: ES cells and somatic cells

BiCell Scientific’s gene targeting vector synthesis service makes targeting vectors to suit a variety of gene editing applications in ES cells as well as in somatic cells including loss-of-gene function, gain-of-gain function, insertion of genetic materials and replacement with human DNA sequences.

Gene targeting can be used to generate knockout, knockin or conditional alleles. The basic targeting vector contains 5′ and 3′ arms of sequence homologous to the gene being targeted and positive/negative selection markers used to disrupt gene function and/or to identify positive cell clones that have integrated targeting vector DNA following transfection.

A targeting vector needs to contain a sufficient length of homology, both 5′ and 3′ to the target gene, in order to permit efficient homologous recombination. While the length of the arms can vary, the homology of the two arms should be at least 5-8 kb in total and the shorter arm no smaller than of 1 kb in length. Targeting frequency may be enhanced when the DNA sequence of the arms is isogenic to the strain from which the ES or somatic cell line was derived.

Positive selection is necessary to identify the targeted cells that have integrated the targeting vector following transfection. The most common positive selection marker is the neomycin resistant (Neo) gene. Negative selection can help distinguish clones that incorporate targeting vector DNA via homologous recombination from those in which the DNA integrates by random insertion. Common negative selection cassettes include HSV-tk or PGK-DTA, which is placed outside the regions of homology in the targeting vector. These markers are lost when the DNA integrates by homologous recombination but retained if inserted randomly. Retention of these markers results in cell death.

BiCell Scientific’s gene targeting vector synthesis service includes genomic DNA synthesis, molecular cloning, targeting sequence assembly, and Sanger sequencing for validation.

  • Gene synthesis of 5′ and 3′ arms of genomic DNA sequences.
  • Molecular cloning of 5′ and 3′ arms into pJ12-PGK-neo-FRT-DTA vector.
  • Molecular cloning of targeting vector carrying intended mutation or insertion (KI projects only).
  • Sanger sequencing of the targeting vector.

Customers will receive the following deliverables at the end of each project:

  • 1 μg of fully assembled targeting vector
  • Sanger sequencing results

For Research Use Only. Not for use in clinical diagnostics or therapeutics.

Additional information

Service type

KO, KI-point mutation, KI-reporter, Floxed-allele, Humanized-allele


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"I am really impressed with your approach. We tried multiple times previously to create monoclonal and polyclonal antibodies to claudin-2 and MLCK1. We have had limited success generating polyclonals and no success generating monoclonals. You have generated outstanding monoclonals to both. I look forward to continuing to work with you."

Jerrold R. Turner, M.D., Ph.D.

Brigham and Women’s Hospital | Harvard Medical School

"The polyclonal antibody you generated for KIAA0408 is stunning! KIAA0408 is a novel cilium molecule that has never been studied. So, clearly there will be a lot of demand for it as we have discovered a very interesting finding and the story will be published in a high impact journal. I am strongly inclined to generate monoclonal antibody for this protein too and we should think about patenting it."

Univ.-Prof. Jay Gopalakrishnan PhD

Heinrich-Heine-Universität | Universitätsklinikum Düsseldorf

"Your ARL13B antibody works beautifully!!! We’re so happy to have a cilia-specific antibody made in rat! I can send you high resolution images to be posted on your website."

Julie Craft Van De Weghe, PhD

School of Medicine | University of Washington

"The assay is a homophilic interaction mediated cell adhesion on purified protein (in this case, immobilized purified Pcdhga9 to Pcdhga9 expressed on cell surface). Compared to control, cell adhesion is reduced in the presence of Pcdhga9 monoclonal antibody supernatants!"

Divyesh Joshi, PhD

School of Medicine | Yale University

"The rabbit hybridoma supernatants of anti-APOBEC3 project are tested positive by ELISA, and we are very happy about it! We previously tried a company, Abclone. Their Project "A" has immune response that is <10,000 titer in antiserum, which would explain why there is no positive mAb after fusion. Their project "B" didn't have any immune response in rabbit."

Harshita B Gupta, PhD.

School of Medicine | UT Health San Antonio

"We have tested anti mouse T cell antiserum samples from both rabbits you sent to us.

They worked very well! Thank you!"

Victoria Gorbacheva, PhD.

School of Medicine | Cleveland Clinic

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