Introducing examples of the use of double knockout and gene knock-in (in mice)!
Here are some examples of the use of the genome editing tool "GenomeCraft." In the case of "double knockout (zebrafish)," Cas9 protein was mixed with sgRNA targeting two genes or GENOME CRAFT Type CT, and injected into zebrafish embryos to observe the development. One day after fertilization, abnormalities in heart formation stained with GFP were observed, and two days later, abnormalities in eye pigment formation were noted. Additionally, there are examples of use in "in vitro digestion assay," "in vivo genome editing," and "gene knock-in (mice)." [In vitro digestion assay use case] ■ Experiment details - Cas9 protein was mixed with sgRNA or crRNA + tracrRNA, along with substrate DNA fragments, to conduct DNA cleavage experiments. ■ Results: A shift in the band was confirmed, indicating that both sgRNA and crRNA + tracrRNA exhibited good DNA cleavage activity. *For more details, please refer to the PDF materials or feel free to contact us.
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【Other Use Cases】 ＜In vivo Genome Editing＞ ■Experiment Details - Mixed Cas9 protein with sgRNA or crRNA + tracrRNA, and injected into zebrafish embryos. ■Results: Multiple bands indicating the presence of mutations were observed for both sgRNA and crRNA + tracrRNA, confirming that it is a highly efficient mutation introduction tool. ＜Gene Knock-in (Mouse)＞ ■Experiment Details - Mixed Cas9 protein with sgRNA targeting the Actb gene or GENOME CRAFT Type CT, along with a targeting donor vector, and injected into mouse fertilized eggs to attempt the creation of knock-in mice. ■Results: Out of 11 mice, 5 knock-in mice were obtained, confirming that genes can be inserted accurately and efficiently at the intended location in the genome. *For more details, please refer to the PDF document or feel free to contact us.
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【Usage Examples】 ■In vitro digestion assay ■In vivo Genome Editing ■Double knockout (zebrafish) ■Gene knock-in (mouse) *For more details, please refer to the PDF document or feel free to contact us.
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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.