Article
Chemistry, Multidisciplinary
Yoo Kyung Kang, Juhee Lee, San Hae Im, Joo Hoon Lee, Juhee Jeong, Duk Ki Kim, Seung Yun Yang, Keehoon Jung, Sang-Gyu Kim, Hyun Jung Chung
Summary: A Cas9 conjugate complex system was developed to induce efficient HDR editing with minimal carrier material. The Cas9-LP complexes showed lower cytotoxicity compared to conventional lipofectamine formulations and achieved efficient base correction of the RFP gene in HEK293T cells.
JOURNAL OF INDUSTRIAL AND ENGINEERING CHEMISTRY
(2021)
Article
Chemistry, Analytical
Alina Passreiter, Nana Naumann, Andreas Thomas, Nicolas Grogna, Philippe Delahaut, Mario Thevis
Summary: The CRISPR/Cas toolkit is a frequently used gene editing technique, but it can also be used illegally for performance enhancement in sports. This study presents a promising strategy for the direct detection of illicit CRISPR/Cas methods in doping control samples using the SHERLOCK system. The method showed specificity and high sensitivity, and was able to detect the presence of CRISPR/Cas traces in serum samples. The proof-of-concept study in a mouse model also demonstrated the potential applicability of the test strategy to authentic doping control samples in the future.
Article
Chemistry, Applied
Toru Taharabaru, Takuya Kihara, Airi Obata, Risako Onodera, Yuting Wen, Jun Li, Keiichi Motoyama, Taishi Higashi
Summary: This study reports the structural fine-tuning and structure-property relationship of multistep transformable poly-rotaxanes (PRXs) for more efficient Cas9 ribonucleoprotein (RNP) delivery. Among various PRXs, PRX derivatives with specific structural properties exhibited the highest genome-editing efficacy and intracellular dynamics control. Furthermore, ligand-modified-8-cyclodextrin improved the cellular uptake and genome-editing effects of the optimized PRX/Cas9 RNP in target cells.
CARBOHYDRATE POLYMERS
(2024)
Article
Nanoscience & Nanotechnology
Min Zhu, Xiuxiu Wang, Ruosen Xie, Yuyuan Wang, Xianghui Xu, Jacobus Burger, Shaoqin Gong
Summary: This study presents a lipopeptide-based nanoparticle (LNP) that can efficiently deliver CRISPR Cas9/sgRNA ribonucleoprotein (RNP) for genome editing both in vitro and in vivo.
ACS APPLIED MATERIALS & INTERFACES
(2023)
Article
Cell Biology
Xingang Yao, Pin Lyu, Kyung Yoo, Manish Kumar Yadav, Ravi Singh, Anthony Atala, Baisong Lu
Summary: A mechanism to actively enrich RNPs into EVs using RNA aptamer and aptamer-binding protein interactions has been described, resulting in efficient genome editing and transient expression. This system allows for multiplex genome editing targeting multiple loci and can deliver RNPs active in vivo. The data demonstrate that this approach could enhance genome editing efficiency and safety.
JOURNAL OF EXTRACELLULAR VESICLES
(2021)
Article
Biochemistry & Molecular Biology
Shirin R. Modarai, Sambee Kanda, Kevin Bloh, Lynn M. Opdenaker, Eric B. Kmiec
Summary: The study revealed that the efficiency and specificity of CRISPR are influenced by multiple factors, including the position of the cleavage site and variability among patient samples. Results showed a unique genetic profile generated based on attempts to correct the SCD single base mutation along the beta globin gene.
Review
Cell Biology
Joseph Andrew Whitley, Houjian Cai
Summary: Clustered regularly interspaced palindromic repeats (CRISPR) is a gene editing tool with great therapeutic potential. Extracellular vesicles (EVs) have emerged as a promising therapeutic delivery vehicle for CRISPR RNP encapsulation. Different strategies have been identified to encapsulate CRISPR RNP into EVs based on genetic fusion, protein tethering, and sgRNA-coupled encapsulation. Incorporating targeting moieties to EVs membrane surface can increase delivery efficiency. However, there are still limitations preventing the use of these strategies in clinical trials.
JOURNAL OF EXTRACELLULAR VESICLES
(2023)
Review
Virology
Dmitriy Mazurov, Lama Ramadan, Natalia Kruglova
Summary: Rapid progress has been made in gene editing using CRISPR/Cas, but the clinical utility is limited due to delivery difficulties and potential off-target effects. Using CRISPR as a ribonucleoprotein (RNP) complex can reduce exposure time and off-target activity. Current delivery systems focus on packaging and delivering CRISPR/Cas RNPs using retro/lentiviral particles and exosomes. The mechanism of viral and exosomal particle formation, release, and entry helps to understand RNP packaging and uncoating. This review also discusses the passive loading of RNPs into exosomes and the mechanisms involved in particle fusion, RNP release, and transportation inside target cells, which can significantly influence the editing efficiency.
Article
Biotechnology & Applied Microbiology
Lu Yu, Meili Xiao, Zhihua Zhu, Yinmei Wang, Zhihua Zhou, Pingping Wang, Gen Zou
Summary: An efficient genome-editing system based on in vitro assembled CRISPR/Cas9 gRNA ribonucleoprotein complexes was developed in C. purpurea. Three target genes were successfully knocked out using this system, resulting in mutants with different phenotypes. This study is of great significance for elucidating the EAS biosynthetic pathway and future basic and applied research on Claviceps purpurea.
SYNTHETIC AND SYSTEMS BIOTECHNOLOGY
(2022)
Article
Biochemical Research Methods
Stephan Riesenberg, Philipp Kanis, Dominik Macak, Damian Wollny, Dorothee Duesterhoeft, Johannes Kowalewski, Nelly Helmbrecht, Tomislav Maricic, Svante Paeaebo
Summary: The study introduces a high-precision method, HDRobust, for repairing DNA double-stranded breaks through homology-directed repair (HDR). By transiently inhibiting nonhomologous end joining and microhomology-mediated end joining, HDRobust achieved point mutations by HDR in up to 93% of chromosomes. Insertions, deletions, rearrangements, and unintended changes at other genomic sites were largely eliminated. The efficient correction of pathogenic mutations in cells derived from patients with anemia, sickle cell disease, and thrombophilia was validated.
Review
Multidisciplinary Sciences
Joy Y. Wang, Jennifer A. Doudna
Summary: The combination of CRISPR genome editing technology and advances in computing and imaging capabilities has brought about a new era in which genetic diseases and individual disease susceptibilities can be predicted and acted upon. This technology has also revolutionized agricultural research and plant breeding by enabling quick identification and alteration of genes responsible for plant traits. In this Review, we discuss the current state of CRISPR-mediated genetic manipulation in human cells, animals, and plants, highlighting successes, challenges, and future prospects.
Article
Biochemistry & Molecular Biology
Trygve A. H. Stromsnes, Sebastian E. E. Schmidke, Mitra Azad, Oyvind Singstad, Idun M. M. Gronsberg, Roy A. A. Dalmo, Arinze S. S. Okoli
Summary: Atlantic salmon is one of the most profitable farmed fish species globally. However, its production growth is threatened by various problems. In this study, gene loci of A. salmon were efficiently edited using CRISPR/Cas technology, and edited cells were enriched and expanded through flow cytometer-activated cell sorting and clonal isolation. This research provides new insights for gene editing of salmonid cells.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Biotechnology & Applied Microbiology
Philipp Reautschnig, Nicolai Wahn, Jacqueline Wettengel, Annika E. Schulz, Ngadhnjim Latifi, Paul Vogel, Tae-Won Kang, Laura S. Pfeiffer, Christine Zarges, Ulrike Naumann, Lars Zender, Jin Billy Li, Thorsten Stafforst
Summary: RNA base editing is a promising alternative to genome editing, and the use of CLUSTER guide RNAs can enhance editing efficiency and precision. The multivalent binding of CLUSTER gRNAs to target messenger RNAs enables editing in various cell lines without bystander effects.
NATURE BIOTECHNOLOGY
(2022)
Review
Biochemistry & Molecular Biology
Hamid Mansouri Khosravi, Michael F. Jantsch
Summary: RNA editing by cytosine and adenosine deaminases changes the identity of edited bases, potentially affecting the coding potential of RNA. Nucleotide deaminases have gained attention for their recoding potential in correcting genetic mutations. RNA editing events are transient, reducing the risk of long-lasting side effects, while some RNA-based therapeutics have been FDA approved for targeting multiple cells or organs to restore genetic function.
Article
Hematology
Suk See De Ravin, Julie Brault, Ronald J. Meis, Siyuan Liu, Linhong Li, Mara Pavel-Dinu, Cicera R. Lazzarotto, Taylor Liu, Sherry M. Koontz, Uimook Choi, Colin L. Sweeney, Narda Theobald, GaHyun Lee, Aaron B. Clark, Sandra S. Burkett, Benjamin P. Kleinstiver, Matthew H. Porteus, Shengdar Tsai, Douglas B. Kuhns, Gary A. Dahl, Stephen Headey, Xiaolin Wu, Harry L. Malech
Summary: Inhibition of p53-binding protein 1 (53BP1) significantly increased long-term homology-directed repair efficiency, leading to highly efficient long-term correction of X-CGD CD34(+) cells.