Review
Biochemistry & Molecular Biology
Chin-Kai Chuang, Wei-Ming Lin
Summary: Cas9-sgRNA is a key tool for genome editing, with its simplicity, unique RNA-guided DNA recognition activity, and independent nuclease activities. Different variants of Cas9, in combination with sgRNA, can achieve various effects on target site breakage. Additionally, engineered enzyme activities can be introduced using Cas9 derivatives, and methods for transient expression and biosafety have been developed to address off-target effects.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Biochemical Research Methods
Yicheng Long, Thomas R. Cech
Summary: The use of CRISPR genome editing has greatly advanced mutagenesis studies in human iPSCs, enabling precise gene manipulation for studying physiological functions, disease modeling, drug screening, and therapeutic development. However, comprehensive strategic designs are necessary to consider potential side effects of CRISPR editing. This study presents novel and highly efficient strategies for introducing mutations in human iPSCs, including step-by-step protocols and examples of successful application in different cell types.
Review
Genetics & Heredity
Chao Li, Wen Chu, Rafaqat Ali Gill, Shifei Sang, Yuqin Shi, Xuezhi Hu, Yuting Yang, Qamar U. Zaman, Baohong Zhang
Summary: In the past decade, significant progress has been made in developing versatile CRISPR/Cas nucleases and precise genome editors. Computer-assisted tools have played a crucial role in designing sgRNA and analyzing CRISPR/Cas-induced mutations. This review provides insights into the different features of CRISPR/Cas tools, the usage of computer-assisted tools, and the limitations and suggestions for further optimization.
GENOMICS PROTEOMICS & BIOINFORMATICS
(2023)
Review
Plant Sciences
Guoning Zhu, Hongliang Zhu
Summary: This article reviews the modifications made to gene-editing systems in recent years and their applications. Modified gene-editing systems such as base editors and prime editors have been developed to enhance the precision of genome engineering. The targeting of mitochondrial genomes and RNA using CRISPR systems has also been explored. Future directions and challenges of modified gene-editing systems are discussed.
FRONTIERS IN PLANT SCIENCE
(2022)
Review
Plant Sciences
A. Giovannini, M. Laura, B. Nesi, M. Savona, T. Cardi
Summary: This review discusses the main genes underlying commercial traits in cut flower species and the possibility of using genome editing techniques to create novel variations and phenotypes. The focus is on promoting flowering and flower longevity, as well as creating novelty in flower structure, color range, and fragrances in ornamental plant breeding. The review also highlights the importance of detailed information about genome and gene structure, as well as improved protocols for efficient delivery of editing reagents for successful implementation of genome editing techniques in elite ornamental crops.
PLANT CELL REPORTS
(2021)
Review
Biology
Alpana Joshi, Seo-Yeon Yang, Hyung-Geun Song, Jiho Min, Ji-Hoon Lee
Summary: Plants are affected by various environmental stresses, which have a negative impact on their growth and development and limit crop productivity. Developing cultivars resistant to abiotic stresses is essential to meet the needs of the growing world population and ensure food security. With the availability of genetic databases and advancements in genome editing techniques, it is now possible to edit target genes with precision and create new opportunities for crop improvement. The CRISPR-Cas system is a powerful genome editing method that allows the development of improved cultivars with resistance to abiotic stresses. This review discusses the current advances in understanding the molecular mechanisms of abiotic stress responses in plants and the application of CRISPR-Cas system genome editing technology for developing crop resilience against drought, salinity, temperature, heavy metals, and herbicides.
Article
Biochemistry & Molecular Biology
Keke Xia, Dengwei Zhang, Xiaojing Xu, Guangyu Liu, Yong Yang, Zhiyong Chen, Xiaojuan Wang, Guo-Qiang Zhang, Hai-Xi Sun, Ying Gu
Summary: This study established a fast and convenient Phalaenopsis protoplast technology for the identification of functional genome editing tools. Two multiplex genome editing systems, PTG-Cas9-HPG and RMC-Cpf1-HPG, were developed and evaluated in Phalaenopsis. The results showed that both systems are functional in Phalaenopsis, and the Super promoter can significantly improve the editing efficiency of the RMC-Cpf1-HPG system. This research provides a valuable foundation for developing diverse and efficient genome editing toolkits and facilitating precise breeding of orchids.
Article
Plant Sciences
Yannick Fierlej, Nathanael M. A. Jacquier, Loic Guille, Jeremy Just, Emilie Montes, Christelle Richard, Jeanne Loue-Manifel, Nathalie Depege-Fargeix, Antoine Gaillard, Thomas Widiez, Peter M. Rogowsky
Summary: In this study, a maize protoplast system coupled with NGS analysis and a novel bioinformatics pipeline was established to evaluate the efficiency of CRISPR/Cas9 tools. The results showed a correlation between mutation types observed in protoplasts and those obtained after stable transformation. Additionally, factors influencing genome editing efficiency were identified, and phenotypic analysis was conducted on base-edited maize plants.
FRONTIERS IN PLANT SCIENCE
(2022)
Review
Microbiology
Qingzhuo Wang, Jiao Zhang, Naief H. Al Makishah, Xiaoman Sun, Zhiqiang Wen, Yu Jiang, Sheng Yang
Summary: Corynebacterium glutamicum is considered a promising biological platform, but faces challenges in genetic manipulation. Recent research shows continuous improvement in genetic manipulation techniques, with CRISPR/Cas-based gene editing tools revolutionizing the field.
FRONTIERS IN MICROBIOLOGY
(2021)
Review
Cell Biology
Marianna Karwacka, Marta Olejniczak
Summary: Polyglutamine diseases are late-onset progressive neurological disorders caused by CAG repeat expansions. The advancement of new gene-editing technologies, particularly the CRISPR-Cas9 technique, has provided valuable tools for studying these diseases.
Review
Genetics & Heredity
Muntazir Mushtaq, Aejaz Ahmad Dar, Milan Skalicky, Anshika Tyagi, Nancy Bhagat, Umer Basu, Basharat Ahmad Bhat, Abbu Zaid, Sajad Ali, Tanvir-Ul-Hassan Dar, Gyanendra Kumar Rai, Shabir Hussain Wani, Muhammad Habib-Ur-Rahman, Vaclav Hejnak, Pavla Vachova, Marian Brestic, Arzu Cig, Fatih Cig, Murat Erman, Ayman EL Sabagh
Summary: Genome-editing technology, particularly the transgene-free approach, has great potential in achieving global food security. Recent developments in CRISPR-based tools for crop improvement include the development of transgene-free genome plants, compatible nucleases, and safe delivery vehicles. Despite the promising advancements, challenges remain in efficient delivery of CRISPR reagents, precision genome editing, and navigating government policies and public acceptance.
Review
Biochemistry & Molecular Biology
Juliet Schreurs, Claudia Sacchetto, Robin M. W. Colpaert, Libero Vitiello, Alessandra Rampazzo, Martina Calore
Summary: In the past two decades, genome editing has emerged as a powerful tool for modeling and treating diseases, with the CRISPR/Cas9 system becoming the most common tool for both disease modeling and therapeutic studies. Novel tools like base editing and prime editing have further expanded the range and accuracy of achievable genomic modifications.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Review
Biochemistry & Molecular Biology
Fangning Zhang, Ting Xiang Neik, William J. W. Thomas, Jacqueline Batley
Summary: CRISPR-Cas is the most prevalent and promising genome editing tool, which plays a crucial role in ensuring global food security and achieving sustainable agricultural development.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Biochemistry & Molecular Biology
Rahul Mahadev Shelake, Dibyajyoti Pramanik, Jae-Yean Kim
Summary: Rapid assessment of CRISPR/Cas-based genome editing tools and components is crucial for successful gene editing applications in various organisms. This study presents a platform for in vivo investigation of CRISPR-BE components in Escherichia coli, enabling base editing without lethal effects on cell survival.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Review
Pharmacology & Pharmacy
Devan L. Puhl, Divya Mohanraj, Derek W. Nelson, Ryan J. Gilbert
Summary: Electrospun fibers are a versatile biomaterial platform for regenerative medicine, with the potential to be used for localized and tunable drug or gene delivery. However, current gene delivery strategies face challenges in clinical applications, which can be overcome by using electrospun fibers for genetic material delivery.
ADVANCED DRUG DELIVERY REVIEWS
(2022)