干细胞基因 z-bo 2020-08-13 556

　　Researchers from the Sloan-Kettering Institute and others in the United States have created a pluripotent stem cell genome editing platform: iCRISPR. This platform can quickly and efficiently knock out genes in stem cells, and it can also be used in the process of stem cell differentiation. Perform stage-specific gene knockout, which will shine in the research of complex pathology of human diseases.

　　Human pluripotent stem cells (hPSCs) can not only be used in clinical regeneration research applications, but also serve as a unique platform for analyzing complex traits and characteristics, elucidating the genes and molecular pathways behind them. In order to achieve this goal, scientists have developed a variety of genetic manipulation methods, but these methods still have various problems. We need fast and manipulable biological methods.

　　In this article, the researchers used CRISPR and TALEN, two of the most concerned genome editing technologies, to develop a human pluripotent stem cell genome editing platform. Researchers call this platform iCRISPR.

　　iCRISPR can be used in the study of gene function loss to quickly and efficiently knock out alleles in human pluripotent stem cells. It can also be used for some precise disease models to perform homozygous pluripotent stem cell knockouts through specific nucleotide transformations. except.

　　Through further experiments, the researchers verified the effectiveness of the one-step method of double and triple knockout hPSC cell lines, and also proved that the stage-specific induction of gene knockout can be performed during the differentiation of pluripotent stem cells, which is useful for developmental biology research. It means a lot.

　　The researchers pointed out that the iCRISPR platform is particularly suitable for analyzing complex genetic interactions and pleiotropic gene functions in human disease research, which will help high-throughput genetic analysis of human pluripotent stem cells.

　　In addition to this study, researchers from the University of California, San Francisco last year proposed a similar name: CRISPRi. They found that when Cas9 lacking endonuclease activity is co-expressed with a guide RNA, a DNA is produced. Recognize complexes that can specifically interfere with transcription elongation, RNA polymerase binding, or transcription factor binding.

　　The researchers developed this CRISPRi system, which can effectively inhibit the expression of targeted genes in E. coli without off-target effects. And with CRISPRi, multiple target genes can be suppressed at the same time, and this effect is also reversible. The researchers also proved that the system is also suitable for gene expression suppression in mammalian cells.

　　Researchers from the Sloan-Kettering Institute and others in the United States have created a pluripotent stem cell genome editing platform: iCRISPR. This platform can quickly and efficiently knock out genes in stem cells, and it can also be used in the process of stem cell differentiation. Perform stage-specific gene knockout, which will shine in the research of complex pathology of human diseases. Related articles were published in the "Cell Stem Cell" magazine on June 12, 2014.

　　Human pluripotent stem cells (hPSCs) can not only be used in clinical regeneration research applications, but also serve as a unique platform for analyzing complex traits and characteristics, elucidating the genes and molecular pathways behind them. In order to achieve this goal, scientists have developed a variety of genetic manipulation methods, but these methods still have various problems. We need fast and manipulable biological methods.

　　In this article, the researchers used CRISPR and TALEN, two of the most concerned genome editing technologies, to develop a human pluripotent stem cell genome editing platform. Researchers call this platform iCRISPR.

　　iCRISPR can be used in the study of gene function loss to quickly and efficiently knock out alleles in human pluripotent stem cells. It can also be used for some precise disease models to perform homozygous pluripotent stem cell knockouts through specific nucleotide transformations. except.

　　Through further experiments, the researchers verified the effectiveness of the one-step method of double and triple knockout hPSC cell lines, and also proved that the stage-specific induction of gene knockout can be performed during the differentiation of pluripotent stem cells, which is useful for developmental biology research. It means a lot.

　　The researchers pointed out that the iCRISPR platform is particularly suitable for analyzing complex genetic interactions and pleiotropic gene functions in human disease research, which will help high-throughput genetic analysis of human pluripotent stem cells.

　　In addition to this study, researchers from the University of California, San Francisco last year proposed a similar name: CRISPRi. They found that when Cas9 lacking endonuclease activity is co-expressed with a guide RNA, a DNA is produced. Recognize complexes that can specifically interfere with transcription elongation, RNA polymerase binding, or transcription factor binding.

　　The researchers developed this CRISPRi system, which can effectively inhibit the expression of targeted genes in E. coli without off-target effects. And with CRISPRi, multiple target genes can be suppressed at the same time, and this effect is also reversible. The researchers also proved that the system is also suitable for gene expression suppression in mammalian cells.