Gene Knock-out Services

CRISPR genes belong to a special DNA repeat sequences family that is ubiquitous in bacterial and archaeological genome. As a "genetic weapon" for defense against foreign genetic material, CRISPR genes can identify and cut foreign DNA to silence their expression. Given its precise targeting, CRISPR system has been developed as a high-efficiency gene editing tool.

crispr gene knock-out

Biologics International Corp (BIC) offers gene knock-out services. We can knock-out any gene of interest in a cell’s genome. Using the target gene sequence provided by the client, we can provide cost-effective gene knockout service, periodic progress updates, and transparent service to solve various client concerns.

Key Features

  • High success rate and low non-specific target rate: multi gRNA design and verification, with non-specific target analysis and testing services
  • Rapid delivery with full transparency: delivery knockout cell lines in ~3 months, periodic progress report, and transparency during the whole process
  • One-stop service: We focus on studies of genes and recombinant proteins. We also provide one-stop service from gRNA design to protein detection (optional)

Service Process

  • Cell preparation:
  • Based on the cell type required, we prepare cells two weeks in advance to maintain their high activity.

  • Identification of target knockout gene:
  • We then identify the coding sequence (CDS) region of the gene and analyze the corresponding genomic structure to determine the exon part of CDS. Based on the nature of genes, candidate knockout sites are selected and then identified.

  • gRNA design and vector construction:
  • Three to five (3–5) groups of gRNA will be designed, and corresponding plasmids will be constructed while also integrating Cas9.

  • Cell transfection and obtaining monoclonal cells:
  • Constructed plasmids are transfected into host cell to generate a cell pool. We will then sequence cells in the cell pool to verify knock out effects.

  • Screening for positive clones:
  • Using limited dilution method, transfected cells are plated on 96-well plate for monoclonal screening. Positive monoclonal cells will be amplified, sequenced, identified, and stored. Finally, a cell line is successfully constructed.

  • Stability test (to be updated):
  • Mycoplasma detection:
  • Quantitative polymerase chain reaction and Western Blot.

  • Cell line freezing and delivery
Need more information? Please do not hesitate to get in touch.

Contact Us

phone+1 (317) 703-0614
fax +1 (855) 427-1516