共查询到20条相似文献,搜索用时 15 毫秒
1.
Maryam Mehravar Abolfazl Shirazi Mahboobeh Nazari Mehdi Banan 《Developmental biology》2019,445(2):156-162
The CRISPR/Cas9 system is a rapid, simple, and often extremely efficient gene editing method. This method has been used in a variety of organisms and cell types over the past several years. However, using this technology for generating gene-edited animals involves a number of obstacles. One such obstacle is mosaicism, which is common in founder animals. This is especially the case when the CRISPR/Cas9 system is used in embryos. Here we review the pros and cons of mosaic mutations of gene-edited animals caused by using the CRISPR/Cas9 system in embryos. Furthermore, we will discuss the mechanisms underlying mosaic mutations resulting from the CRISPR/Cas9 system, as well as the possible strategies for reducing mosaicism. By developing ways to overcome mosaic mutations when using CRISPR/Cas9, genotyping for germline gene disruptions should become more reliable. This achievement will pave the way for using the CRISPR technology in the research and clinical applications where mosaicism is an issue. 相似文献
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CRISPR(clustered regularly interspaced short palindromic repeats)/Cas9(CRISPR-associated proteins)作为一种新型基因组编辑技术,为解释疾病的发生机制和治疗疾病提供了新方法。来自Ⅱ型原核CRISPR系统的CRISPR/Cas9能够通过单链向导RNA(single guide RNA, sgRNA)将Cas9核酸酶靶定到特定的基因组序列发挥作用。已经被成功用来进行基因编辑构建疾病模型,以进行相关领域的功能研究和疾病的治疗。CRISPR/Cas9技术正在迅速的应用于生物医学研究的各个领域,包括心血管领域,它促进了人们对电生理、心肌病、心律失常以及其他心血管疾病的更多了解,已经创建了靶向很多基因的细胞和动物模型,为新一类疗法打开了大门。本综述介绍了CRISPR/Cas9的作用原理、优点和局限性,以及在心血管疾病中的应用进展。 相似文献
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Wang Bang Lv Xiujuan Wang Yufei Wang Zhibo Liu Qi Lu Bin Liu Yong Gu Feng 《中国科学:生命科学英文版》2021,64(9):1463-1472
Genetic manipulation of mitochondrial DNA(mtDNA) could be harnessed for deciphering the gene function of mitochondria; it also acts as a promising approach for the therapeutic correction of pathogenic mutation in mtDNA. However, there is still a lack of direct evidence showing the edited mutagenesis within human mtDNA by clustered regularly interspaced short palindromic repeats-associated protein 9(CRISPR/Cas9). Here, using engineered CRISPR/Cas9, we observed numerous insertion/deletion(InDel) events at several mtDNA microhomologous regions, which were triggered specifically by double-strand break(DSB)lesions within mtDNA. InDel mutagenesis was significantly improved by sgRNA multiplexing and a DSB repair inhibitor,iniparib, demonstrating the evidence of rewiring DSB repair status to manipulate mtDNA using CRISPR/Cas9. These findings would provide novel insights into mtDNA mutagenesis and mitochondrial gene therapy for diseases involving pathogenic mtDNA. 相似文献
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Geupil Jang Sangyool Lee Tae Young Um Sun Hyun Chang Han Yong Lee Pil Joong Chung Ju-Kon Kim Yang Do Choi 《Plant biotechnology reports》2016,10(6):425-435
The CRISPR/Cas9 technology is useful for genome editing to generate targeted mutants. Based on this genome editing technology, it was attempted to generate the rice mutant which lacks JAZ9 activity to understand its function in stress response. The sequence of guide RNA for the recognition of target site was obtained from CRISPR-PLANT website (http://www.genome.arizona.edu/crispr) to minimize off-target effect and was recombined into the CRISPR/Cas9 binary vector pRGEB31. Embryonic calli regenerated from the mature seeds (O. sativa L. cv. Nakdong) were co-cultivated with transformed Agrobacterium tumefaciens LBA4404, and 26 individual transgenic plants were obtained through the hygromycin selection process. Nucleotide sequence analysis showed that most of T0 plants carried both edited and unedited wt sequence of JAZ9, suggesting genetic chimerism of T0 plants. Even though 2 individual lines carried homozygous mutation on JAZ9, they were also chimeric due to biallelic mutation. The relative ratio between edited and unedited wt sequence was variable among individual lines. Expression level of Cas9 is correlated with the frequency of genome editing frequency. In some plants, the enrichment ratio changed along with developmental stage. The nucleotide sequence analysis revealed that Cas9-mediated A/T addition occurred at -3 nucleotide position from protospacer adjacent motif (PAM), whereas G addition at -5 nucleotide position from the PAM. Further analysis of T1 transgenic plants showed that the genome editing patterns were similar between T0 plants and their T1 sibling plants. These suggested that earlier selection of T0 plants with homozygous mutation is an efficient way to obtain homozygous mutants in T1 generation. 相似文献
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Permyakova Natalya V. Sidorchuk Yury V. Marenkova Tatyana V. Khozeeva Sofya A. Kuznetsov Vitaly V. Zagorskaya Alla A. Rozov Sergei M. Deineko Elena V. 《Molecular biology reports》2019,46(6):5735-5743
Molecular Biology Reports - Targeted genome editing using CRISPR/Cas9 is a promising technology successfully verified in various plant species; however, it has hardly been used in plant cell... 相似文献
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Akhmetova E. A. Golyshev V. M. Vokhtantcev I. P. Meschaninova M. I. Venyaminova A. G. Novopashina D. S. 《Russian Journal of Bioorganic Chemistry》2021,47(2):496-504
Russian Journal of Bioorganic Chemistry - A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable... 相似文献
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Kankan Wang Xiaochun Tang Zicong Xie Xiaodong Zou Mengjing Li Hongming Yuan Nannan Guo Hongsheng Ouyang Huping Jiao Daxin Pang 《Transgenic research》2017,26(6):799-805
CRISPR/Cas9 has emerged as one of the most popular genome editing tools due to its simple design and high efficiency in multiple species. Myostatin (MSTN) negatively regulates skeletal muscle growth and mutations in myostatin cause double-muscled phenotype in various animals. Here, we generated myostatin mutation in Erhualian pigs using a combination of CRISPR/Cas9 and somatic cell nuclear transfer. The protein level of myostatin precursor decreased dramatically in mutant cloned piglets. Unlike myostatin knockout Landrace, which often encountered health issues and died shortly after birth, Erhualian pigs harboring homozygous mutations were viable. Moreover, myostatin knockout Erhualian pigs exhibited partial double-muscled phenotype such as prominent muscular protrusion, wider back and hip compared with wild-type piglets. Genome editing in Chinese indigenous pig breeds thus holds great promise not only for improving growth performance, but also for protecting endangered genetic resources. 相似文献
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《中国科学:生命科学英文版》2017,(5)
正Dear Editor,The CRISPR/Cas9(clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9)system is revolutionizing genome editing due to its high efficiency,low cost,design simplicity and versatility.However,introduction of a point mutation at a desired position remains a great challenge in plant genome engineering.Currently,point mutation in plants was achieved by incorporating a 相似文献
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Hydrobiologia - Reverse genetics approaches are critical for uncovering complex biological processes and genetic engineering. Clustered regularly interspaced short palindromic repeats... 相似文献
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Jinzhi Duan Guangqing Lu Zhan Xie Mingliang Lou Jiao Luo Lei Guo Yu Zhang 《Cell research》2014,24(8):1009-1012
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<正>Geminiviruses are a group of circular single-stranded DNA viruses that constitute the largest family of plant viruses.Many diseases resulting from geminivirus infections, such as maize streak disease, cassava mosaic disease, tomato yellow leaf curl disease and cotton leaf curl diseases, cause significant problems in terms of economic losses, posing a serious threat to global crop productivity (Hanley-Bowdoin et al., 2013; Yang et al., 2019). As obligate intracellular 相似文献
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Zhi Miao Qian Li Jian Zhao Peng Wang Lei Wang Hong-Peng He Nan Wang Hao Zhou Tong-Cun Zhang Xue-Gang Luo 《Biotechnology letters》2018,40(8):1209-1218
Objectives
To establish stable infliximab-expressing Chinese hamster ovary (CHO) cells with high tolerance to serum-free culture.Results
Bcl-2 antagonist/killer 1 (BAK1), which is a key mediator of the apoptosis pathway, was disrupted, and infliximab, which is a broadly used monoclonal antibody for the treatment of rheumatoid arthritis and other autoimmune diseases, was incorporated into the BAK1 locus of the CHO chromosome using the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas genome-editing technique. The activating effects of serum starvation on BAK1 and cytochrome C (CytC) were suppressed in the genome-edited cells, and the ability of the cells to resist the serum starvation-induced loss of mitochondrial membrane potential and apoptosis was increased, as indicated by the results of polymerase chain reaction (PCR), flow cytometry, enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography (HPLC) analysis. In addition, during subsequent passages, infliximab could be stably produced in the genome-edited CHO cells, and the recombinant antibody could effectively antagonize the cytotoxic effect of tumor necrosis factor α (TNFα).Conclusions
A CHO cell line capable of stably expressing infliximab and adapting to serum-free culture was constructed. This work lays the foundation for the development of infliximab biosimilars.17.
Chihiro Kadooka Masaaki Yamaguchi Kayu Okutsu Yumiko Yoshizaki Kazunori Takamine Takuya Katayama 《Bioscience, biotechnology, and biochemistry》2020,84(10):2179-2183
ABSTRACT We developed an approach to genome editing of the white koji fungus, Aspergillus luchuensis mut. kawachii using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system. Co-transformation of AMA1-based Cas9 and gRNA expression plasmids achieved efficient gene knockout in A. kawachii. The plasmids were easily lost when selective pressure was removed, allowing for successive rounds of genome editing. 相似文献
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CRISPR/Cas9 genome editing in wheat 总被引:1,自引:0,他引:1
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《遗传学报》2016,(2)
The clustered regularly interspaced short palindromic repeats(CRISPR)/Cas9 system, a simple and efficient tool for genome editing, has experienced rapid progress in its technology and applicability in the past two years. Here, we review the recent advances in CRISPR/Cas9 technology and the ways that have been adopted to expand our capacity for precise genome manipulation. First, we introduce the mechanism of CRISPR/Cas9, including its biochemical and structural implications. Second, we highlight the latest improvements in the CRISPR/Cas9 system, especially Cas9 protein modifications for customization. Third, we review its current applications, in which the versatile CRISPR/Cas9 system was employed to edit the genome, epigenome, or RNA of various organisms. Although CRISPR/Cas9 allows convenient genome editing accompanied by many benefits, we should not ignore the significant ethical and biosafety concerns that it raises. Finally, we discuss the prospective applications and challenges of several promising techniques adapted from CRISPR/Cas9. 相似文献
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《中国科学:生命科学英文版》2017,(5)
正Dear Editor,Proper development of plant roots is critical for primary physiological functions,including water and nutrient absorption and uptake,physical support,and carbohydrate storage(Zhang et al.,2010).Crop root systems act as the key organ for sensing and response to abiotic and biotic stresses.Previous studies of crop root system development suggest that increased lateral root formation(LRF)could stimulate 相似文献