Destruction of Being "Lifted": A Study on Klara and the Sun from the Perspective of Bioethics / 中国医学伦理学

Klara and the Sun is the first work published by Kazuo Ishiguro after he won the Nobel Prize for Literature. Its publication attracted widespread attention for he turned his attention to the gene-editing technology. By telling the story of Clara, an artificial friend (AF) designed to accompany "lifted" children, the author revealed the ethical problems caused by the application of gene-editing technology to "lifted" children. From the theoretical perspective of bioethics and its basic principles, this paper interpreted the ethical issues of gene-editing technology from three aspects that: "lifted" technology, "lifted" technology implementer, and "lifted" technology implementation consequence, and analyzed Kazuo Ishiguro’s technical ethics proposition.

Legal Regulatory Approach of Germline Cell Gene Editing / 中国医学伦理学

Germline cell gene editing is a potentially revolutionary technology that can be used to treat and prevent many genetic diseases, such as cystic fibrosis and hemophilia. Although this technology has aroused widespread discussion in the medical community and the public, its legal regulatory approach has not yet been fully determined. By summarizing and studying the experience of relevant foreign laws and regulations, this paper analyzed and summarized the current legal situation in China, and explored three ethical topics of social equality, individual subjectivity, and intergenerational rights. Combining the existing legal norms and relevant ethical considerations, the legal regulatory approach for germline cell gene editing was proposed in view of the weak operability of the law, the blurred boundaries of the legislation, and the insufficiently perfect ethical review mechanism. To better ensure that the technology can benefit mankind on the premise of eliminating risks, the following suggestions were put forward: a specialized law should be formulated based on clear legislative principles to adjust and guide the improvement of the ethical review mechanism of germline cell gene editing. It is hoped that promoting the progress of germline cell gene editing technology through in-depth research, and providing useful reference for the formulation of relevant laws and ethical policies.

Application of CRISPR/Cas9 genome editing technology in the research of eye diseases / 中华实验眼科杂志

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) is an adaptive immune system against invasive viruses and exogenous DNA,which is developed by bacteria and archaer during long-term evolution.With advances in technology,researchers have found that CRISPR/ Cas9 system can precisely edit the genomes of eukaryotic cells through insertion,replacement or deletion of target genes.Using CRISPR/Cas9 genome editing technology,researchers found that overexpression of paired box gene 6 (PAX6) in cornea can cause congenital corneal epithelial damage;this technology promoted the research on the pathogenic mechanism of keratin 12 (KRT12) mutation in Meesmann corneal epithelial dystrophy;it has also built congenital cataract animal models by knocking out the G JA8 gene and αA lens gene,which is beneficial to the application of the diagnosis and pathological analysis of congenital cataract.In addition,the researchers have used CRISPR/Cas9 genome editing technology to confirm the correlation of RHOS334 mutated RHO allele,P23H mutated RHO gene,Y347X mutated Pde6b gene,and mutant RP9 allele with retinitis pigmentosa.The application of this technology has provided evidence to support the association of KCNJ13 gene and mutant CEP290 gene with Leber congenital amaurosis.CRISPR/Cas9 can provide target spot of intraocular neovascular diseases the targeted therapy by editing of VEGFR2 gene and TXNIP gene,and it also playse an important role in the study of pathogenic genes and establishment of animal models for proliferative vitreoretinopathy and retinoblastoma.In this review,we introduced the evolutionary history,the molecular characteristics and the mechanism of CRISPR/Cas9,and summarized its current research advances in eye diseases.

Discussion on Ethical Review Question of CRISPR/Cas9 Gene Editing Technology in Clinical Research / 中国医学伦理学

Objective:To explore the ethical review questions of CRISPR/Cas9 gene editing technology caused in clinical research,and thus to provide a reference for matters needing attention of the ethical review involved in this technology in clinical.Methods:This paper summarized the ethical problems of CRISPR/Cas9 gene editing technology at home and abroad,analyzed the reasons and put forward some suggestions for the application of new technology in line with China's national conditions.Results:It should allow CRISPR/Cas9 gene editing technology to be applied in somatic cell gene therapy,forbidden for genital gene therapy and not considered to enhance.Since CRISPR/Cas9 lack clear subject of responsibility ethics,it brings security,conflict of rights and social equality issues.The measures that need to be taken include the strengthening of cultural communication,the formation of ethic forms of gene editing technology,the establishment of independent ethical review body at national's level,the improvement of legal norms,the formulating of technical standards and ethical principles and the major support to gene editing research field at the national level.Conclusion:In view of the potential clinical application of CRISPR/Cas9,our country should progressively restrictively develop the embryonic gene editing techniques from the prohibition.The ethics committee is responsible for the ethical review and supervision of clinical research.Members of the ethics committee and ethical staff should strengthen the study of new knowledge,strictly docking policies and regulations from accepting the clinical research projects involving CRISPR / Cas9 to ethical review,and thus to ensure the effectively review the ethical problems of genetic editing technology project.

Construction of Nedd4 knockout BMDM cell line by CRISPR/Cas9 technology / 军事医学

Objective To construct Nedd4 knockout bone marrow-derived macrophages(BMDM) cell line by CRISPR/Cas9 technology and to provide an effective tool for studying the function and mechanism of Nedd4 in macrophage.Methods First,three high-grade sgRNAs targeting Nedd4 gene exons were screened using the online tool before synthesized sgRNAs were inserted into the PX330 plasmid respectively.Secondly,the recombinant plasmids were transferred into BMDM cells and monoclonal cells were obtained by limiting dilution method.The protein levels of NEDD4 in monoclonal cells were detected by Western blotting.Finally,the DNA sequence of the monoclonal cells was confirmed by sequence analysis.Results One Nedd4 knockout BMDM cell line was obtained.The sequencing result showed that the Nedd4 gene had 16bp deletion mutation in this cell line.Conclusion The Nedd4 knockout BMDM macrophage cell line constructed by CRISPR/Cas9 technology will be a useful tool for studying the function and mechanism of Nedd4 in BMDM cells.

Application of CRISPR/Cas9 in plant biology

The CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins) system was first identified in bacteria and archaea and can degrade exogenous substrates. It was developed as a gene editing technology in 2013. Over the subsequent years, it has received extensive attention owing to its easy manipulation, high efficiency, and wide application in gene mutation and transcriptional regulation in mammals and plants. The process of CRISPR/Cas is optimized constantly and its application has also expanded dramatically. Therefore, CRISPR/Cas is considered a revolutionary technology in plant biology. Here, we introduce the mechanism of the type II CRISPR/Cas called CRISPR/Cas9, update its recent advances in various applications in plants, and discuss its future prospects to provide an argument for its use in the study of medicinal plants.

CRISPR/Cas9-mediated Editing of Human β-globin Gene in Human Cells: A Commentary on the Research Ethics.

Abstract: Recently, Chinese researchers published the results of their research using a gene-editing technology on abnormal human zygotes. The research team believes this research has prospective clinical application, viz., for gene therapy for β-thalassemia, a white blood cell disorder, and plan to persist with further studies, despite technical problems in this experiment. The research has elicited international criticism from both scientific and bioethics domains, because it innovates beyond the current global consensus against human germ line modification. This paper comments on some ethical issues presented by the research report and concludes that, under present circumstances, the Chinese research team did not meet a standard of scientific responsibility.