Review
Cell Biology
Gemma Llargues-Sistac, Laia Bonjoch, Sergi Castellvi-Bel
Summary: The use of next-generation sequencing (NGS) technologies has greatly advanced our understanding of the mutational landscape of complex human diseases such as cancer. Haploid human cell models, such as the HAP1 cell line, have emerged as valuable tools for functional gene studies, especially in combination with CRISPR-Cas9 gene editing technology. This review explores the recent applications of the HAP1 cell line in functional genetic studies and high-throughput genetic screens, highlighting its potential to enhance our understanding of gene function and the genetic basis of human diseases identified through NGS technologies, and its implications for clinical practice and patient care.
FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY
(2023)
Article
Microbiology
Xueli Zhang, Chaohui Zhang, Caijiao Liang, Bizhou Li, Fanmei Meng, Yuncan Ai
Summary: Bacteriophages, the most abundant organisms in the biosphere, have been sequenced extensively. However, the study of bacteriophage functional genomics has been hindered by a lack of effective research methods. This study designed a phage genome editing platform based on the CRISPR-Cas9 system, and successfully achieved gene editing in Vibrio natriegens phage TT4P2. This platform has the potential to advance research on phage gene diversity and accelerate the development of phage synthetic biology and nanotechnology.
MICROBIOLOGY SPECTRUM
(2022)
Review
Biochemistry & Molecular Biology
Yulin Mu, Chengxiao Zhang, Taihua Li, Feng-Jie Jin, Yun-Ju Sung, Hee-Mock Oh, Hyung-Gwan Lee, Long Jin
Summary: Lactobacillus, important in food production and probiotics, can benefit from CRISPR/Cas9-based genome editing to improve strain efficiency and achieve traceless genome modification.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Biochemical Research Methods
Wenxiu Yin, Hanhua Hu
Summary: In this study, a short transformation period genetic transformation system was established in Chaetoceros muelleri using CRISPR/Cas9 technology. The editing efficiency was up to 86%, providing a molecular tool for the study of basic biology in C. muelleri and its synthetic biology applications.
ACS SYNTHETIC BIOLOGY
(2023)
Article
Veterinary Sciences
Chun-Yu Liu, Ming Jin, Hao Guo, Hong-Zhe Zhao, Li-Na Hou, Yang Yang, Yong-Jun Wen, Feng-Xue Wang
Summary: Bovine herpesvirus type I (BoHV-1) is a highly pathogenic virus that causes respiratory diseases in cattle, resulting in significant economic losses to the cattle industry worldwide. Researchers successfully knocked out the gE gene of BoHV-1 using the CRISPR/Cas9 system and observed interesting phenomena. This study provides a foundation for future research on BoHV-1 and vaccine development.
VETERINARY SCIENCES
(2022)
Article
Biochemistry & Molecular Biology
Azamat V. Karginov, Marina G. Tarutina, Anastasia R. Lapteva, Maria D. Pakhomova, Artur A. Galliamov, Sergey Y. Filkin, Alexey N. Fedorov, Michael O. Agaphonov
Summary: In this work, an improved plasmid vector set for CRISPR-Cas9 genome editing in methylotrophic yeasts is presented. The vector set includes a plasmid encoding Cas9 with a nuclear localization signal and plasmids with a scaffold for the single guide RNA (sgRNA). Construction of sgRNA gene for a specific target sequence only requires the insertion of a 24 bp oligonucleotide duplex into the scaffold. The functionality of this vector set has been demonstrated in multiple methylotrophic yeast species.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Biochemical Research Methods
Stefan J. Tekel, Nicholas Brookhouser, Kylie Standage-Beier, Xiao Wang, David A. Brafman
Summary: The introduction of transient reporters of editing enrichment (TREE) has enabled highly efficient single-base editing of human cells using a transient episomal fluorescent reporter, allowing for rapid generation of clonal editing efficiencies exceeding 80% in biallelic or multiplexed edited isogenic human pluripotent stem cell lines within approximately 3-4 weeks.
Review
Chemistry, Physical
Noshad Peyravian, Maziar Malekzadeh Kebria, Jafar Kiani, Peiman Brouki Milan, Masoud Mozafari
Summary: This review focuses on the recent advancements of microfluidic systems to deliver CRISPR/Cas systems in clinical and therapy investigations, highlighting the limitations of conventional delivery tools and the advantages of microfluidic systems in in vitro delivery.
Article
Immunology
Nicoletta Schibeci Natoli Scialli, Barbara Colitti, Luigi Bertolotti, Giulia Pezzoni, Eugenio Martignani, Maverick Melega, Emiliana Brocchi, Sergio Rosati
Summary: The CRISPR/Cas9 system has greatly improved genome editing and gene expression technologies, allowing for stable and high-yield production of recombinant proteins. This study successfully utilized the system to edit a hybridoma cell line for stable expression of proteins of interest, demonstrating its potential for specific applications in infectious disease diagnostics.
VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY
(2021)
Review
Biochemistry & Molecular Biology
Pascal D. Vos, Aleksandra Filipovska, Oliver Rackham
Summary: The discovery and widespread use of CRISPR-Cas9 has greatly impacted biological research. However, Cas9 has certain limitations that need improvement for its use in biotechnology. Scientists have developed promising Cas9 variants through protein engineering campaigns, which can improve and expand the scope and fidelity of CRISPR applications.
BIOCHEMICAL SOCIETY TRANSACTIONS
(2022)
Review
Agriculture, Dairy & Animal Science
Julia Popova, Victoria Bets, Elena Kozhevnikova
Summary: Genome editing has practical applications in farm animals, improving production traits, economic value, and disease resistance. It also has potential in biomedical research and drug production, as well as xenograft donors. Recent advancements in site-specific nucleases and embryological delivery methods have revolutionized transgenesis, providing efficient and reliable tools for genome engineering in agriculture.
Review
Pharmacology & Pharmacy
Jie Li, Joachim Justad Raise, Maomao He, Riddha Das, Niren Murthy
Summary: Genome-editing tools like CRISPR have revolutionized various fields, but the low delivery efficiency of genome editing proteins in vivo remains a major challenge, with the most successful strategy currently involving potentially immunogenic viral vectors.
ADVANCED DRUG DELIVERY REVIEWS
(2021)
Article
Biochemistry & Molecular Biology
Patrick Ferreira, Altino Branco Choupina
Summary: In recent years, the number of tools and techniques for precise genome editing has increased significantly. The CRISPR/Cas9 technology has gained much attention due to its simplicity, affordability, and effectiveness. It has the potential for various applications in gene editing, including functional genomics, cell model development, targeted therapies, and improvement of agricultural and environmental species.
MOLECULAR BIOLOGY REPORTS
(2022)
Review
Pharmacology & Pharmacy
Siwei Chen, Deng Chen, Bin Liu, Hidde J. Haisma
Summary: CRISPR/Cas9-mediated genome engineering has extensive applications in basic biology, biotechnology, and medicine. However, the low gene modification efficiency and uncontrollable prolonged Cas9 activity hinder its further use. Researchers have explored small molecules with clinical potential to precisely modulate CRISPR/Cas9 genome editing activity.
DRUG DISCOVERY TODAY
(2022)
Article
Biochemistry & Molecular Biology
Xing Chen, Qing Tan, Qingyang Lyu, Chengxuan Yu, Na Jiang, Jianqiang Li, Laixin Luo
Summary: The study presents an efficient unmarked CRISPR/Cas9-mediated gene editing system for Clavibacter michiganensis, enabling site-directed mutagenesis and complete gene knockout. The inclusion of codA::upp in the transformation vector allowed for counterselection of unmarked mutants, making the system more convenient and reusable for editing multiple genes.
MOLECULAR PLANT-MICROBE INTERACTIONS
(2022)