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  6. Random Integration Analysis Service using the RAISING Method

Random Integration Analysis Service using the RAISING Method

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last updated:Apr 22, 2021

ファスマック
ファスマック
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Developed through joint research with the National Institute of Infectious Diseases. Achieves determination of the insertion site of foreign DNA in a short time, with high sensitivity and low cost.

The "RAISING method" is a technique for detecting the insertion sites of foreign DNA in the genome. It allows for rapid and reproducible measurement results with a small amount of DNA. It can identify insertion sites of viral genome insertions, viral vector insertions, searches for fusion genes that could be drug discovery targets, and off-target knock-ins in genome editing, among others. 【Features】 - Analysis can be arranged according to the purpose and target. - Multiple insertion sites can be detected in a single analysis. - High reproducibility of results is achieved through proprietary technology. 【Analysis Examples】 - HTLV-1 (Human T-cell Leukemia Virus) Clonality Analysis Leveraging the characteristics of the RAISING method, which can quickly detect with a small amount of DNA, we achieved high detection sensitivity even with a proviral load of 0.032%. - Off-target Detection of Knock-ins in Genome Editing For knock-in mice created using CRISPR/Cas, we conducted detection of knock-in efficiency and off-target sites by combining with NGS. *For details on examples and services, please refer to the materials available for download.

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rais_method_about1.png

Random Integration Analysis Service using the RAISING Method

rais_method_about1.png
rais_method_about1.png
  • Related Link - http://fasmac.co.jp/rais_method_about

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■ We offer two service plans: "Sanger Sequencing" and "NGS" ▼ Sanger Sequencing Plan - Usage examples: Viral genomes, analysis of the clonality of fusion genes, etc. - Deliverables: Waveform data (ab1 files), sequence data (txt files) ▼ Next Generation Sequencing (NGS) Plan - Usage examples: Off-target detection of knock-ins using genome editing or viral vectors, etc. - Deliverables: Quality-value annotated Miseq output sequences (fastq files) 【Other Applied Technologies Introduction】 "Antibody Sequence Sequencing Analysis" By obtaining the variable sequences of antibodies, we support the production of recombinant antibodies. The variable sequences of mouse IgG genes expressed in hybridomas are amplified using the 5' RACE method or RT-PCR, followed by DNA cloning in E. coli and sequencing analysis. "Long Chain RNA Synthesis" We synthesize long chain RNA using in vitro transcription methods. Please provide the desired sequences for various applications such as RT-PCR amplification control RNA, gRNA for genome editing, small RNA, mRNA preparation, RNA aptamers, etc.

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Applications/Examples of results

【Usage Examples】 Determine the positions where foreign DNA is randomly inserted into the host genome, including the target location. ◆ Virus genome insertion sites ◆ Virus vector insertion sites ◆ Search for fusion genes that could be drug discovery targets ◆ Detection of off-target knock-ins in genome editing 【Analysis Examples】 ■ HTLV-1 Clonality Analysis ■ Detection of off-target knock-ins in genome editing 【Regarding Sample Requests】 Please provide purified genomic DNA at a concentration of 200 ng/μl or more, with a minimum volume of 10 μl per sample. *If the sample amount is small, please consult with us. *Quality checks and RNase treatment are mandatory. *For more details, please refer to the PDF document or feel free to contact us.

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DNA random integration analysis

DNA random integration analysis

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Genome editing tool 'GENOME CRAFT RNA'

Genome editing tool 'GENOME CRAFT RNA'

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Next-generation sequencing analysis

Next-generation sequencing analysis

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Fasmac Inc. Company Profile

Fasmac Inc. Company Profile

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Fasmac Inc. "Technical Introduction Materials - New Added Value to Existing Platforms -"

Fasmac Inc. "Technical Introduction Materials - New Added Value to Existing Platforms -"

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Poster presentation on HTLV-1 clonal analysis technology.

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There will be a poster presentation on the HTLV-1 clonality analysis technology jointly developed by Phasmac and the National Institute of Infectious Diseases at the HTLV-1 Society Academic Meeting to be held next month. November 6 (Saturday) 17:30-17:50 P-18 "Examination of the newly developed HTLV-1 clonality analysis technology as an ATL risk assessment method" Additionally, this has been selected for the Excellent Poster Presentation award. 7th Japan HTLV-1 Society Academic Meeting Date: November 5 (Friday) to 7 (Sunday), 2021 Venue: Kumamoto Castle Hall

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[Case Study] Determination of Insertion Sites for Animal Cultured Cells and Plant Cells

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Using the Agrobacterium method, we inserted T-DNA taglines into Arabidopsis thaliana and performed the RAISING method on mouse cultured cells with retrovirus vectors. We confirmed the insertion site sequences using the Sanger method and mapped the decoded sequences obtained from next-generation sequencing analysis to the genome sequence. As a result, it was confirmed that this method is effective for determining insertion positions in both animal cultured cells and plant cells.

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6th Annual Meeting of the Japanese Society for Genome Editing Poster Presentation

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We will present a poster at the 6th Annual Meeting of the Japanese Society for Genome Editing. Dates: June 16 (Wednesday) to June 18 (Friday), 2021 Format: Online Our presentation details are as follows: June 17 (Thursday) 17:10 - 17:50 P-11B Detection Method for Foreign DNA Insertion Sites Using Random Integration Analysis We will introduce a method for detecting off-target effects of knock-ins in genome editing. Please take a look.

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ファスマック

ファスマック

Pharmaceuticals and Biotechnology

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In recent years, advancements in molecular biology have led to remarkable progress in research such as the determination of the complete genome sequences of organisms and subsequent gene function analysis. Along with these technological advancements, molecular biological techniques such as nucleotide sequence analysis, real-time PCR, and DNA chips have become widely utilized in the field of food analysis. Since its establishment in 2001, Fasmak Co., Ltd. has been developing technologies for the "Japanese Standard Analysis Method" for genetically modified foods and food allergens in collaboration with related agencies of the Ministry of Agriculture, Forestry and Fisheries and the Ministry of Health, Labour and Welfare. The developed testing technologies are provided not only in Japan but also in the United States, China, and other countries. Additionally, Fasmak has been actively engaged in international standardization activities for "food inspection methods using molecular biological techniques" since its establishment, and its technical capabilities are internationally recognized. Furthermore, in partnership with Eurofins Scientific, one of the world's largest testing companies, Fasmak is advancing the introduction of the high technical capabilities possessed by the Eurofins Group. Moving forward, Fasmak will continue to provide world-class new testing technologies to everyone.

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