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CRISPR/Cas9 Platform provides a base editing service for generating a point mutation. By this service, one base can be converted to another: C to T, or A to G. With years of experience in genome editing, our scientists are dedicated to bring customer high-quality service with the latest technology.

CRISPR/Cas9 system is one of the most common used gene-editing tools, which was first described as an immune defense system. Considering its easy-to-use, simple-to-design and multiplexed engineering, CRISPR/Cas9 is widely used for genome engineering in microbes, mammalian cells/models and plants. However, the inefficiency of precise base editing and off-target activities remains. To get rid of this situation, base editing is developed as a new genome editing technology, which can be widely used in mammalian cells, plants, microbes and model organisms. Base editing enables the irreversible conversion of a specific DNA base into another at a targeted genomic locus, for example conversing C to T, or A to G. Unlike other genome-editing tools, base editing can be achieved without double-strand breaks. When introducing a point mutation at a target locus, base editing is more efficient than traditional genome editing techniques. Since many genetic diseases arise from point mutations, base editing has important applications in disease research.

CRISPR/Cas9 Platform provides one-stop gene knockin cell line generation service, including reporter gene knockin, tag knockin and fragment knockin. This service ranges from design of gRNA, vector construction, cell transfection and selection, to single clone sequencing and cell expansion. Our talented scientists are expert in knockin cell line generation by using CRISPR system. At CRISPR/Cas9 platformCB, you can expect to achieve with satisfying services and products.

Knockin cell line, which inserted a certain fragment in a specific locus, is a useful tool for research. When a reporter gene or tag is inserted at the specific locus, this cell line can be used to study endogenous expression level of targeted gene. With the help of CRISPR/Cas9 system, it become easier to generate a custom knockin cell line. Cas9 nuclease targets to different loci of genome through sgRNA with different sequences and then the enzyme introduces double strand break. At the presence of template, DNA double strand break can be repaired via HDR pathway. By this method, promoter, tag or gene can be inserted into genome.

CRISPR/Cas9 Platform provides a point mutation cell line generation service for various cell lines, such as tumor cell lines and stem cell lines. As a provider of genome editing service, CRISPR/Cas9 PlatformCB focuses on using the latest technique to serve customers. With years of experience, our scientist team is expert in generating knock-in stable cell lines with CRISPR. At CRISPR/Cas9 PlatformCB, all projects are done by well-trained staff with excellent platform under the guide of talented scientists. We provide customers with reliable results and products.

Gene mutation is an important inducer for many diseases. With CRISPR/Cas9 system, it becomes possible to correct the mutated site in living cells. This technique is based on two factors, guide RNA and Cas9 nuclease. Guide RNA alters its sequence for different target sites, while Cas9 functions as a nuclease cutting double strand DNA. At the presence of template, homology directed repair pathway is activated for reconstructing the target site. Due to its versatility, CRISPR system is widely used all over the world and promotes the development of many research fields.

CRISPR/Cas9 Platform provides one stop custom knock-in cell line generation service, including point mutation and gene insertion. With non-stop exploration of technology, we have developed an excellent platform for gene editing. Based on the platform, our expert staff has succeeded in dozens of knock-in cell line generation projects, including stem cells, tumor cells and even difficult-to-handle cells. At CRISPR/Cas9 PlatformCB, a full cell line generation service is offered from sgRNA design/construction to final cell line generation/verification.

CRISPR system is a novel tool for precise gene editing. Currently, the most commonly used system is derived from Streptococcus pyogenes (Sp), which consists of a Cas9 nuclease and a guide RNA. SpCas9 will induce a DNA double-stranded break after recruited to specific site by sgRNA. The subsequent homologous recombination repair will generate a knock-in or point mutation phenotype in the presence of a recombination donor. Thus, the HDR pathway are exploited to facilitate correction of diseased genes, insertion of epitope tags or fluorescent reporters, and overexpression of genes of interest in a site-specific manner.

Engineered nucleases, which can introduce targeted DNA double-stranded breaks into the genome of living cells, are widely used as genome-editing tools. However, they may introduce undesired off-target mutations as well, potentially leading to unintended effects in research and clinical applications. Since indel mutations can not be easily detected by anchored PCR amplification of an expected sequence, sensitive and unbiased genome-wide methods are developed to understand the genome locations and frequency of nuclease-induced DSB activity.

As one of the most popular methods, circularization for in vitro reporting of cleavage effects by sequencing (CIRCLE-Seq) is a novel, highly sensitive, sequencing-efficient strategy to identify CRISPR/Cas9 genome-wide off-target mutations. This assay can be practiced using widely accessible next-generation sequencing technology since it does not require reference genome sequences. Thus, CIRCLE-Seq is applied for off-target profiling in organisms lacking full genomic sequence. Comparing to Digenome-seq, CIRCLE-seq shows lower background by enrichment of nuclease-cleaved genomic DNA before sequencing. Additionally, CIRCLE-seq, as a sensitive method, can detect off-target mutations that occur with frequencies below ~0.1% in nuclease-treated cell populations.

Creative Biogene CRISPR/Cas9 Platform provides a GUIDE-Seq service to assess off-target effect of CRISPR/Cas9. At CRISPR/Cas9 PlatformCB, all projects are performed by skillful staff under the guide of experienced scientists. We are dedicated to assisting you and paving the way for your successful research with reliable services and products.

Since CRISPR/Cas9 technology was applied for genome engineering, it brings a revolutionary development of genome-editing technology. CRISPR system makes it easy and efficient to make targets cuts to genome. However, it may induce cleave off-target DNA targets at low frequencies when applied for genome editing. To overcome this limitation, various methods have developed to detect these unexpected cleavages by scientists. Genome-wide Unbiased Identifications of DSBs Evaluated by Sequencing (GUIDE-Seq) is one of cell-based assays for off-target sites detection. This assay is an approach for global detection of DNA double-stranded breaks introduced by RGNs (CRISPR RNA-guided nucleases) and potentially other nucleases. It is highly sensitive and can detect off-target sites that occur at a frequency of 0.1% in a cell population. GUIDE-Seq is a recommended method to detect the putative off-target cleavage sites.

CRISPR/Cas9 Platform provides an off-target prediction and analysis service for CRISPR/Cas9-mediated gene editing. With years of experiences, our scientists are expert in application of CRSIPR/Cas9 system. Our scientist team will provide reliable and professional off-target testing services for our clients. As a leader in gene editing, CRISPR/Cas9 PlatformCB offers trusted analysis results with high quality in a competitive price.

Cas9, which is a nuclease, can be recruited to the specific site by guide RNA and then cleaves DNA. Similarly to other nucleases, Cas9 can cleave off-target DNA targets in the genome at reduced frequencies. For application of Cas9, it is essential to consider ways to minimize the degree of off-target cleavage and to detect the presence of off-target cleavage. Though many tips have been developed to minimize the risk of off-target cleavage, off target mutations is still a major concern about CRISPR/Cas9-mediated genome editing in some instance. These mutations should be carefully monitored, especially when using CRISPR/Cas9 for therapeutic purposes. However, off-target analyses of the CRISPR/Cas9 system have been very challenging, particularly when performed directly in cells. With the wide application of next generation sequence, different sequencing strategies have been developed to identify the off-target mutations, such as whole genome sequencing, GUIDE-seq, Digenome-seq.

CRISPR/Cas9 Platform delivers high-quality CRISPR/Cas9-mediated genome editing service for our clients. Our microbial genome editing service are backed by extensive experience, top researchers and biosecure facilities. CRISPR/Cas9 PlatformCB deploys a powerful and flexible gene editing platform to help you every step of the way with innovative bioanalytical solutions in your genome editing workflow.

Homologous recombination has been broadly applied in genome editing of prokaryotes with high efficiency and accuracy. However, this way is limited in realizing larger-scale genome editing with large DNA fragments or numerous genes because DNA editing template construction is a relatively complicated procedure1. Derived from the immune system present in bacteria and archaea, CRISPR technology as an efficient genome-scale editing tool has revolutionized conventional genetic engineering methods and unprecedentedly facilitated strain engineering2. Consequently, the CRISPR/Cas9 system has rapidly become a transformative and powerful tool for editing genomes in many organisms.