Article
Plant Sciences
Masahiro Nishihara, Akiko Hirabuchi, Fumina Goto, Yuzo Nishizaki, Shota Uesugi, Aiko Watanabe, Keisuke Tasaki, Rie Washiashi, Nobuhiro Sasaki
Summary: Genetic engineering was used to produce yellow-flowered gentians by introducing betaxanthin pigments. The white-flowered gentians were first created by knocking out the DFR gene, and then the betaxanthin biosynthetic genes were transformed. The resultant gentian plants exhibited vivid yellow flowers with specific expression and accumulation of seven known betaxanthins.
Article
Multidisciplinary Sciences
Zhaohui Zhong, Guanqing Liu, Zhongjie Tang, Shuyue Xiang, Liang Yang, Lan Huang, Yao He, Tingting Fan, Shishi Liu, Xuelian Zheng, Tao Zhang, Yiping Qi, Jian Huang, Yong Zhang
Summary: In this study, a probiotic sourced CRISPR-LrCas9 system with a similar PAM requirement to Cas12a was reported, and its high efficiency in various genome editing applications was demonstrated.
NATURE COMMUNICATIONS
(2023)
Article
Cell Biology
Bo Li, Chunyang Fu, Jiawei Zhou, Fengjiao Hui, Qiongqiong Wang, Fuqiu Wang, Guanying Wang, Zhongping Xu, Lianlian Che, Daojun Yuan, Yanqin Wang, Xianlong Zhang, Shuangxia Jin
Summary: Establishing efficient gene editing techniques in cotton is of great importance for studying gene function. Researchers found that the pBeYDV-Cas9-KO system has higher editing efficiency and specificity when editing the GhCLA1 gene compared to the traditional CRISPR/Cas9 system.
Article
Biochemistry & Molecular Biology
Mohd Farhan Siddiqui, Moon-Moo Kim
Summary: Melanin is a key pigment in skin and hair, with deficiency leading to various disorders. Enhancing melanin production at a genetic level could offer an effective and permanent solution for these disorders, with the gene SIRT7 potentially playing a crucial role in this process.
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE
(2021)
Article
Biochemistry & Molecular Biology
Jianduo Zhang, Jiaxin Xing, Qili Mi, Wenwu Yang, Haiying Xiang, Li Xu, Wanli Zeng, Jin Wang, Lele Deng, Jiarui Jiang, Guangyu Yang, Qian Gao, Xuemei Li
Summary: CRISPR/Cas9 genome-editing technology has revolutionized plant science, showing great potential for crop improvement. By editing the NtPDS gene in tobacco, researchers have demonstrated that incorporating an OsU3-tRNA promoter combination into the CRISPR/Cas9 system can enhance editing efficiency. The optimized tobacco CRISPR/Cas9 system, named pOREU3TR, successfully produced transgene-free and homozygous gene-edited plants with significant differences in plant height and amino acid content. This OsU3-tRNA-sgRNA/AtUb10-Ros1 system provides essential improvements for increasing the efficiency of plant genome editing.
Article
Agriculture, Dairy & Animal Science
Celia Pineiro-Silva, Sergio Navarro-Serna, Ramses Belda-Perez, Joaquin Gadea
Summary: Lipofection treatment, which involves encapsulating the CRISPR/Cas9 system into vesicles and introducing them into cells, can be a simple and inexpensive method for producing genetically modified pigs. It has similar effects on embryo development and mutation rates as the electroporation technique when used in zona-pellucida-intact porcine oocytes. However, increasing the concentration of liposomes-CRISPR/Cas9 system complexes in the media has a detrimental effect on embryo development parameters, possibly indicating a toxic effect.
Article
Medicine, Research & Experimental
Jae Ho Kim, Moon-Moo Kim
Summary: This study aimed to investigate the relationship between melanin production and lipofuscin formation, and found that TYR gene knockout cells exhibited decreased melanin production and increased lipofuscin formation.
Article
Biochemistry & Molecular Biology
Haniel Siqueira Mortagua Walflor, Aline Rodrigues Castro Lucena, Felipe Francisco Tuon, Lia Carolina Soares Medeiros, Helisson Faoro
Summary: Antimicrobial resistance is a major global public health concern and was identified as one of the main causes of death in 2019. This study successfully restored the sensitivity to fosfomycin using a CRISPR-Cas9 system and identified effective guide RNAs for resensitization.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Microbiology
Jianyu Liu, Haiyang Cui, Ruijuan Wang, Zhen Xu, Hailong Yu, Chunyan Song, Huan Lu, Qiaozhen Li, Danrun Xing, Qi Tan, Weiming Sun, Gen Zou, Xiaodong Shang
Summary: This study developed an optimized CRISPR/Cas9 genome-editing method using in vitro assembled ribonucleoprotein complexes (RNPs) in Flammulina filiformis. The method achieved a 100% targeting efficiency and avoided the use of foreign DNA. This research has significant theoretical and practical implications.
Article
Plant Sciences
Junpeng Li, Wenbo Pan, Shuai Zhang, Guojing Ma, Aixia Li, Huawei Zhang, Lijing Liu
Summary: The study found that GRF4-GIF1 and GRF5 can effectively enhance the transformation efficiency of sorghum, and the combination with helper plasmid pVS1-VIR2 further enhances the transformation. In addition, a sorghum CRISPR/Cas9 gene-editing tool was developed based on the GRF4-GIF1/ternary vector system.
Article
Biotechnology & Applied Microbiology
Rajveer Singh, Shivani Chandel, Arijit Ghosh, Anupam Gautam, Daniel H. Huson, V. Ravichandiran, Dipanjan Ghosh
Summary: In the past two decades, yeast has been widely used as a cost-effective biological tool for producing small molecules and biofuels. The application of CRISPR-Cas techniques in yeast genetic and metabolic engineering has revolutionized the field, particularly in targeted chromosomal integration using synthetic donor constructs. This study presents a highly efficient strategy for targeted chromosomal integration and in-frame expression of foreign genes in the yeast genome using CRISPR-Cas9 and homology-dependent recombination (HDR). The optimized methods achieved a 74% efficiency in integrating small constructs at multiple target sites. The study also demonstrated that as few as 15 base pairs of microhomology are sufficient for targeted knock-in with minimal construct concentration. Whole-genome sequencing confirmed the absence of off-target effects. This protocol provides a comprehensive and streamlined approach for integrating essential genes into the yeast genome for synthetic biology and industrial purposes.
Article
Horticulture
Yuxiang Liu, Min Wei, Yumei Liu, Zhiyuan Fang, Yangyong Zhang, Limei Yang, Honghao Lv, Yong Wang, Jialei Ji, Xiaoli Zhang, Zhansheng Li, Fengqing Han
Summary: CRISPR/Cas9 system is utilized for precise gene editing in broccoli, a globally important vegetable, by introducing multiple single-guide RNA-expressing cassettes. The study demonstrates that CRISPR/Cas9 can be a promising tool for gene functional verification and trait improvement in broccoli.
SCIENTIA HORTICULTURAE
(2023)
Article
Multidisciplinary Sciences
Zsolt Bodai, Alena L. Bishop, Valentino M. Gantz, Alexis C. Komor
Summary: Programmable double-strand DNA breaks (DSBs) can be harnessed for precision genome editing through manipulation of the homology-directed repair (HDR) pathway. This study introduces a general strategy called the double tap method, which improves HDR-mediated precision editing efficiency by taking advantage of the reproducible nature of indel sequences. The method utilizes multiple gRNAs, including a primary gRNA that targets the wild-type genomic sequence and one or more secondary gRNAs that target the most common indel sequence(s).
NATURE COMMUNICATIONS
(2022)
Article
Plant Sciences
Wenwen Kong, Mingliang Wang, Lijuan Huang, Feiyan Wu, Jinyuan Tao, Beixin Mo, Yu Yu
Summary: CRISPR/Cas9-dependent genome editing has been successfully applied to various plant species for fundamental studies and crop improvement, enabling random or targeted insertions, deletions, and base changes. To simplify the screening process for edited transformants, a dsRED2 cassette driven by CaMV 35S was introduced into the Cas9 vector, providing a visible protein for easy genetic screening without the need for instruments. Moreover, the use of the WUS promoter to drive the expression of Cas9 improved editing efficiency and enabled homozygous mutagenesis of target genes in Arabidopsis, making this system potentially applicable to other crops.
Article
Biology
Tao Ye, Yangyang Duan, Hayley W. S. Tsang, He Xu, Yuewen Chen, Han Cao, Yu Chen, Amy K. Y. Fu, Nancy Y. Ip
Summary: Researchers demonstrate a simple and precise method to simultaneously generate iPSC lines with different gene dosages using paired Cas9 nickases. As proof-of-concept they apply this method to examining amyloid precursor protein gene dosage effects in an Alzheimer's disease patient-derived iPSC line. Their method could potentially advance what we know about disease mechanisms and assist with gene therapy development.
COMMUNICATIONS BIOLOGY
(2021)
