Introduction
Genome editing is a genetic manipulation technique that can modify DNA sequences at the genome level. The principle of this technology is to construct an artificial endonuclease to cut off DNA at a predetermined genome location, and the cut DNA will produce mutations in the process of being repaired by the DNA repair system in the cell, so as to achieve the purpose of site-specific modification of the genome.
CRISPR is a gene-editing technology for the bacterial immune system that recognizes target cell DN The nucleotide sequence of the target in the segment uses the endonuclease protein to cleave the DNA target sequence to produce a double-stranded DNA break, which will be repaired by the cell repair mechanism in vivo, among which, the repair of non-homologous end-attached nhej will produce a short nucleic acid insertion or deletion, and the gene frameshift mutation brought by it will lead to the early appearance of the stop codon, so as to complete the knockout of the target cell DNA target gene fragment. The Cas9 gRNA gene drum removal system recognizes a specific DNA sequence through a small gRNA segment and cleaves the DNA near the recognition site, so as to achieve site-directed knockout or knock-in of the gene. Since specific gene recognition is accomplished by a small piece of RNA, i.e., gRNA, while the protein components in the system remain unchanged, it is very convenient to use different synthetic or expressed gRNAs to cooperate with Cas9 protein to achieve knockout of different genes.
A mature and stable experimental system for gene interference expression has been established, which can provide you with gene knockout cell line construction services for a variety of different cells (including difficult-to-transfect cells). It has a mature cell experiment service platform, and the service content is rich and detailed. At the same time, it can also provide customers with a variety of technical services including overexpression, silencing and stable cell line construction.
Step 1
1. Screening of stable transfection cell lines.
2 Vector construction.
3 Virus packaging.
4 Cell transfection.
5. Screening of stably transfected cell lines.
6 Delivery (negotiable).
Experiment report. Construct viral vectors.
Gene expression test report of interest.
Knockout cells
Cell selection
1 There must be no mycoplasma contamination.
2 Select cell lines with successful cases.
3. Cells are easy to proliferate, have a high clone formation rate, have no polygenic problems, and do not mutate too much chromosomes.
4 Tumor cell lines.
Scope of application
1. To study the function and expression regulation mechanism of genes.
2. Find a suitable drug target.
Advantages:
l Success rate and fast delivery speed;
l Service-oriented: reasonable, according to the comprehensive analysis of customer needs, to provide customers with satisfactory results;
l Professional technical support, to ensure strong, thoughtful and timely pre-sales and after-sales service support, ready to answer any questions for you;
l Complete upstream and downstream services: with a complete service platform for cells, antibodies, proteins, etc., and one-stop technical services, saving customers' valuable time and money.
l A variety of lentiviral expression vectors with different promoters and tags are available
l Cells are clear and of good quality without bacterial, fungal and mycoplasma contamination
Service process
After the two parties communicate with each other, determine the experimental plan, determine the service requirements, sign the contract, make an advance payment, start the experiment, and deliver the results.