Gene Knock out services
Given its precise targeting, CRISPR system has been developed as a high-efficiency gene editing tool. Wisecorner Laboratories Pvt Ltd 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
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High success rate and low non-specific target rate: multi gRNA design and verification, with off target analysis and testing services
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Rapid delivery with full transparency: delivery knockout cell lines in ~3 months, periodic progress report, and transparency during the whole process
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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
Based on the cell type required, we prepare cells two weeks in advance to maintain their high activity.
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.
Three to five (3–5) groups of gRNA will be designed, and corresponding plasmids will be constructed while also integrating Cas9.
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.
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.
Quantitative polymerase chain reaction and Western Blot.
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Cell preparation:
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Identification of target knockout gene:
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gRNA design and vector construction:
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Cell transfection and obtaining monoclonal cells:
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Screening for positive clones:
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Stability test (to be updated):
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Mycoplasma detection:
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Cell line freezing and delivery
Lipid Nanoparticle Synthesis and Encapsulation Service:
Wisecorner Laboratories Pvt Ltd offers full range of services from mRNA synthesis till its encapsulation using Lipid Nanoparticles followed by its analytical characterization.
Lipid nanoparticles (LNPs) are the most clinically advanced non-viral gene delivery system. Lipid nanoparticles safely and effectively deliver nucleic acids, overcoming a major barrier preventing the development and use of genetic medicines. Genetic medicine has many different applications such as gene editing, rapid vaccine development, immuno-oncology and treatment of rare genetic and undruggable diseases; all of which are usually hindered by nucleic acid delivery inefficiency.
Lipid nanoparticles offer many advantages over previous lipid-based nucleic acid delivery systems including:
• High nucleic acid encapsulation efficiency and potent transfection
• Improved penetration into tissues to deliver therapeutics
• Low cytotoxicity and immunogenicity
These characteristics make lipid nanoparticles excellent candidates for nucleic acid delivery.