One way to overcome the challenges of studying the epigenome systematically is the use of manipulated genome editing tools. This approach is known as epigenome editing and relies on altering gene expression without altering the underlying sequences and is thus truly epigenetic. Epigenome editing systems rely on the targeting capabilities of a genome editing system, but are modified to have the nuclease inactivated or removed so it is incapable of cleaving DNA, but can still reach its target.
The novel functionality is given by a fusion of the altered genome editing system to an effector domain, which is a modular domain that gives the system the power of an epigenome modifier. Thus removing or inactivating the nuclease and adding an effector domain to provide new function confers the modular powers of a swiss army knife and allows for manipulation of the epigenome in several different ways.
One-stop technical services are provided from gene information, cells, sequencing, analysis to construction of cell lines by our company. Our CRISPR genome editing technology applies to targets of any genes and mammalian cells. Professional researchers have plentiful CRISPR genome editing experience, including gRNA design, cell transfection, monoclonal cell culture and passages, solving difficulty in transfection and constructing tumor cell lines. After services are finishes, we will provide gene-edited monoclonal cells and detailed project reports.
1. Choose transfection methods of the specific cell: Try all kinds of methods, including liposome-mediated transfection, electroporation, nucleofection, lentivirus-mediated transduction, et al.
2. Determine antibiotic concentration: Decrease difficulties in selecting positive cells and checking them.
3. Culture monoclonal cells: Determine if monoclonal cells are cultured.
Three gRNAs are designed in common exons of different transcriptional products, generally. Target positions are as far as possible in the first third of the gene, and targets can destroy important domains of the protein and all the alternatively spliced transcripts. Six gRNAs are designed if the effects are not good or the time is short.
Construct vectors or pack lentiviruses
Common Cas9 vectors or lentiviral vectors are selected according to the results of pre-experiments. Vector plasmids and lentiviruses are prepared.
Transfect the cells with vector plasmids or transduct the cells with lentivirus.
Analysize cell pool
Genomic PCR amplicons are sequenced to confirm if genome editing is produced.
Select single clones and bank positive cells
Single positive clones are selected according to the sequence results and these cells are banked.
Present final data
Provide clients with single cell clones and detailed reports.
Generally, cell lines are provided by customers. An additional charge is collected if our company provide them. In addition, 200 tumor cell lines are offered except general cell lines.
Additional single clone: The other single clone are selected from the same cell bank and presented to clients with analysis of target sequences included in final report. Functional identification of single clones: qPCR and western blot of the target protein.
1. Provide real one-stop service from gene synthesis, design of gRNAs and donors, cell transfection, preparing of single clones to sequencing analysis of single clone, only needing customers to present target genes
2. Do not use antibiotic to select cells and fuse tags in target genes(only in gene knock-out project)
3. Possess rich cell culture experiences
4. Do not add antibiotic in cell culture. Ensure aseptic operation and sterile environment strictly.
1. Target gRNA sequences are designed(free of charge), synthesized and cloned into any vectors according to target sequences provided by customers.
2. Provide construction service of stable overexpression cell lines and help you to develop stable cell lines expressing foreign genes.
In order to provide quotation efficiently and accurately, please download and complete the inquiry sheet, then fill out the whole inquiry sheet and email it to us, we will reply to you in time.
The product can only be used for scientific research purposes, NOT be used in any human or clinical experiments, NOT be used to modify any human reproductive system, including the human embryo and reproductive cells.