Review
Pharmacology & Pharmacy
Daniel Allen, Nechama Kalter, Michael Rosenberg, Ayal Hendel
Summary: Genome engineering through targeted nucleases, especially CRISPR-Cas9, has revolutionized gene therapy research and has the potential to treat blood and immune system diseases. CRISPR-Cas9 homology-directed repair (HDR) represents a promising method for site-specific gene insertion or correction. Other methods, like viral gene addition, gene knockout through non-homologous end joining (NHEJ), and base or prime editing, also show promise but have significant limitations for treating genetic immune or blood disorders. This review aims to highlight the transformative benefits of HDR-mediated gene therapy and propose solutions for current challenges, in order to advance HDR-based gene therapy in CD34(+) hematopoietic stem progenitor cells (HSPCs).
Article
Biochemistry & Molecular Biology
Chengpeng Wang, Yang Li, Na Wang, Qin Yu, Yonghong Li, Junping Gao, Xiaofeng Zhou, Nan Ma
Summary: We established a platform for CRISPR/Cas9-mediated gene editing in rose using suspension cells, and successfully edited a gene involved in the ethylene signaling pathway. Our optimized CRISPR/Cas9 system provides a powerful tool for functional genomics, molecular breeding, and synthetic biology in rose.
JOURNAL OF INTEGRATIVE PLANT BIOLOGY
(2023)
Article
Plant Sciences
Jilin Chen, Shaoya Li, Yubing He, Jingying Li, Lanqin Xia
Summary: This article summarizes the latest developments in CRISPR/Cas-mediated homology-directed repair technology for plants, highlighting significant achievements in crop improvement. It describes various strategies to enhance repair efficiency, as well as the challenges and open questions in precision genome editing for both plant biological research and crop improvement.
Article
Genetics & Heredity
Dongjin Kim, Megan Hager, Eleanor Brant, Hikmet Budak
Summary: Genome editing using CRISPR technology in wheat can be more efficient with the use of LbCpf1, compared to Cas9, for enhancing the editing efficiency of target genes. However, Cas9 may induce more off-target mutations than LbCpf1, indicating the potential of CRISPR-LbCpf1 as a powerful genome editing tool for polyploid plants like wheat.
FUNCTIONAL & INTEGRATIVE GENOMICS
(2021)
Article
Biotechnology & Applied Microbiology
Jose Bonafont, Angeles Mencia, Esteban Chacon-Solano, Wai Srifa, Sriram Vaidyanathan, Rosa Romano, Marta Garcia, Rosario Hervas-Salcedo, Laura Ugalde, Blanca Duarte, Matthew H. Porteus, Marcela Del Rio, Fernando Larcher, Rodolfo Murillas
Summary: This study presents a gene-editing approach using CRISPR-Cas9 system to achieve gene correction in different cell types, showing therapeutic potential for RDEB.
Review
Chemistry, Multidisciplinary
Deepak Kumar Sahel, Lalitkumar K. Vora, Aishwarya Saraswat, Saurabh Sharma, Jasmin Monpara, Anisha A. D'Souza, Deepakkumar Mishra, Kamatham Pushpa Tryphena, Satoru Kawakita, Shahid Khan, Mohd Azhar, Dharmendra Kumar Khatri, Ketan Patel, Raghu Raj Singh Thakur
Summary: CRISPR and Cas9, initially discovered as an immune system in bacteria and archaea, are now widely used as biotechnological tools for genetic and nongenetic disease treatment. Clinical trials mainly adopt ex vivo strategies, but in vivo delivery methods for CRISPR components are still under preclinical surveillance.
Article
Genetics & Heredity
Siyu Chen, Zhiquan Liu, Wanhua Xie, Hao Yu, Liangxue Lai, Zhanjun Li
Summary: In this study, two compact CjeCas9 orthologs (Cje2Cas9 and Cje3Cas9) were identified, which can efficiently edit the genome in human cells and have simpler PAM requirements, expanding the targeting scope. It was also found that Cje3Cas9 can successfully disrupt target genes in mice and showed potential for therapeutic applications.
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.
Article
Biology
Zhiquan Liu, Siyu Chen, Wanhua Xie, Hao Yu, Liangxue Lai, Zhanjun Li
Summary: Researchers have revisited and engineered a compact Cas9 orthologue derived from Neisseria cinerea (NcCas9) for efficient genome editing in mammal cells. NcCas9 can recognize a PAM sequence (N4GYAT) that existing Cas9s cannot, and by optimizing its architecture and spacer length, editing efficacy is improved. NcCas9-derived Base editors can efficiently generate base conversions, and six anti-CRISPR proteins were identified as off-switches for NcCas9. NcCas9 successfully generated efficient editing of mouse embryos by microinjection of NcCas9 mRNA and the corresponding sgRNA.
COMMUNICATIONS BIOLOGY
(2022)
Article
Multidisciplinary Sciences
Raed Ibraheim, Phillip W. L. Tai, Aamir Mir, Nida Javeed, Jiaming Wang, Tomas C. Rodriguez, Suk Namkung, Samantha Nelson, Eraj Shafiq Khokhar, Esther Mintzer, Stacy Maitland, Zexiang Chen, Yueying Cao, Emmanouela Tsagkaraki, Scot A. Wolfe, Dan Wang, Athma A. Pai, Wen Xue, Guangping Gao, Erik J. Sontheimer
Summary: This study introduces a single-vector AAV platform for effective genome editing therapy, along with a nanopore-based sequencing platform for quality control. The results demonstrate successful treatment of two genetic disease models in mice using these platforms.
NATURE COMMUNICATIONS
(2021)
Article
Pharmacology & Pharmacy
Lingmin Zhang, Langyu Yang, Jionghua Huang, Sheng Chen, Chuangjia Huang, Yinshan Lin, Ao Shen, ZhouYikang Zheng, Wenfu Zheng, Shunqing Tang
Summary: A novel gene vector ZEBRA was developed for efficient delivery of the CRISPR/Cas9 system, showing cell line dependency with approximately 10-fold higher transduction efficiency in CD44-positive cancer cell lines compared to CD44-negative ones.
ASIAN JOURNAL OF PHARMACEUTICAL SCIENCES
(2022)
Review
Plant Sciences
Yingxin Guo, Guangdong Zhao, Xing Gao, Lin Zhang, Yanan Zhang, Xiaoming Cai, Xuejiao Yuan, Xingqi Guo
Summary: This review provides a direction for crop quality improvement and ideas for further research on the application of CRISPR/Cas9 gene editing technology for crop improvement. It discusses the achievements of CRISPR/Cas9 gene technology in improving crop quality and the challenges and prospects of this technology.
Article
Chemistry, Physical
Juhee Lee, Yoo Kyung Kang, Eonju Oh, Juhee Jeong, San Hae Im, Duk Ki Kim, Haeshin Lee, Sang-Gyu Kim, Keehoon Jung, Hyun Jung Chung
Summary: The study presents a cancer gene therapy strategy based on NanoRNP that efficiently blocks the PD-L1 immune checkpoint and induces an antitumor effect in vivo without the need for combination therapy. In vivo results demonstrate that NanoRNP can induce indels in target cells at high frequencies, significantly suppressing tumor growth.
CHEMISTRY OF MATERIALS
(2022)
Article
Immunology
Huan Qin, Wenliang Zhang, Shiyao Zhang, Yuan Feng, Weihui Xu, Jia Qi, Qian Zhang, Chunxiu Xu, Shanshan Liu, Jia Zhang, Yushuang Lei, Wanqin Liu, Shuyu Feng, Jingjing Wang, Xuefei Fu, Zifen Xu, Ping Li, Kai Yao
Summary: The researchers developed a genome-editing tool called PESpRY, which combines the versatility of prime editors with the unconstrained PAM requirement of the SpRY Cas9 variant. Using this tool, they were able to successfully correct gene mutations in a mouse model of retinitis pigmentosa, leading to substantial restoration of vision.
JOURNAL OF EXPERIMENTAL MEDICINE
(2023)
Article
Biochemistry & Molecular Biology
Xiaoying Zhao, Kunli Qu, Benedetta Curci, Huanming Yang, Lars Bolund, Lin Lin, Yonglun Luo
Summary: Recent progress in CRISPR gene editing tools has expanded the possibilities for treating devastating genetic diseases. In this study, three methods of gene editing (NHBEJ, HDR, and PE) were compared for correcting loss-of-function mutations in Duchenne Muscular Dystrophy. The highest efficiency was achieved with NHBEJ, followed by HDR and PE2. The correction efficiency was increased with the use of PE3. This study demonstrates the potential for highly efficient correction of DMD mutations using CRISPR gene editing.
Article
Hematology
Glenn F. Pierce, K. John Pasi, Donna Coffin, Radoslaw Kaczmarek, David Lillicrap, Johnny Mahlangu, Dawn Rottellini, Thomas Sannie, Alok Srivastava, Thierry VandenDriessche, Alain Weill
Article
Genetics & Heredity
Jennifer Bolleyn, Matthias Rombaut, Nisha Nair, Steven Branson, Anja Heymans, Marinee Chuah, Thierry VandenDriessche, Vera Rogiers, Joery De Kock, Tamara Vanhaecke
Article
Hematology
Nisha Nair, Dries De Wolf, Phuong Anh Nguyen, Quang Hong Pham, Ermira Samara-Kuko, Jeff Landau, Grant E. Blouse, Marinee K. Chuah, Thierry VandenDriessche
Summary: The novel FIX variant CB 2679d-GT showed significantly improved clotting activity and hemostatic correction in hemophilic mice compared to the current standard R338L-Padua variant following AAV-directed gene therapy. This study highlights the potential of CB 2679d-GT to achieve higher FIX activity levels and superior hemostatic efficacy in hemophilia B patients.
Article
Biotechnology & Applied Microbiology
Alberto Malerba, Chiara Sidoli, Ngoc Lu-Nguyen, Shan Herath, Anita Le Heron, Hayder Abdul-Razak, Susan Jarmin, Thierry VandenDriessche, Marinee K. Chuah, George Dickson, Linda Popplewell
Summary: Duchenne muscular dystrophy (DMD) is a genetic disease affecting male individuals, with no cure currently available. Gene therapy, particularly using AAV vectors, shows promise in improving expression levels and relieving symptoms in patients.
HUMAN GENE THERAPY
(2021)
Article
Biotechnology & Applied Microbiology
Sumitava Dastidar, Debanjana Majumdar, Jaitip Tipanee, Kshitiz Singh, Arnaud F. Klein, Denis Furling, Marinee K. Chuah, Thierry VandenDriessche
Summary: CTG repeat expansion is associated with cardiac dysfunction in DM1. Excision of the CTG repeat corrects the aberrant alternate splicing and reverses the abnormal cardiac spliceopathy in DM1. Correction of certain genes is important for cardiac development, maturation, and function.
Article
Multidisciplinary Sciences
Sourav K. Bose, Brandon M. White, Meghana V. Kashyap, Apeksha Dave, Felix R. De Bie, Haiying Li, Kshitiz Singh, Pallavi Menon, Tiankun Wang, Shiva Teerdhala, Vishal Swaminathan, Heather A. Hartman, Sowmya Jayachandran, Prashant Chandrasekaran, Kiran Musunuru, Rajan Jain, David B. Frank, Philip Zoltick, William H. Peranteau
Summary: In utero base editing shows potential in correcting disease-causing mutations before birth, improving survival and alleviating symptoms of diseases like MPSI. This approach highlights the feasibility and efficacy of therapeutic base editing in multiple organs before birth, offering promising treatment options for genetic diseases.
NATURE COMMUNICATIONS
(2021)
Article
Biotechnology & Applied Microbiology
Jaitip Tipanee, Ermira Samara-Kuko, Thierry Gevaert, Marinee K. Chuah, Thierry VandenDriessche
Summary: This study developed a non-viral platform based on Sleeping Beauty transposons and minicircles for genetic modification, allowing efficient expression of CD19-28z.CAR and inactivation of allogeneic TCRs using CRISPR-Cas9. The resulting CAR T cells showed anti-tumor activity against CD19+ tumor cells and induced complete tumor remission in a mouse model, while minimizing TCR alloreactivity and GvHD. This non-viral approach provides an alternative method for generating next-generation CD19-specific CAR T cells, reducing GvHD risk and manufacturing constraints associated with viral vectors.
Editorial Material
Biotechnology & Applied Microbiology
Thierry VandenDriessche, Steven W. Pipe, Glenn F. Pierce, Radoslaw Kaczmarek
Article
Genetics & Heredity
Kshitiz Singh, Brianna M. Quinville, Melissa Mitchell, Zhilin Chen, Jagdeep S. Walia
Summary: Sandhoff disease is a fatal neurodegenerative disorder without curative treatment. The study found that immune system related genes are significantly upregulated in the disease progression, while the expression levels of myelination related genes are not statistically significant. There is an age-dependent increase in the expression of microglial/pro-inflammatory genes, while the expression of myelination related genes slightly decreases or remains unchanged.
Editorial Material
Biotechnology & Applied Microbiology
Thierry VandenDriessche, Marinee K. Chuah
Article
Biochemistry & Molecular Biology
Julieth A. A. Sierra-Delgado, Shibi Likhite, Paula K. K. Bautista, Sergio A. A. Gomez-Ochoa, Luis E. E. Echeverria, Elizabeth Guio, Clara Vargas, Norma C. C. Serrano, Kathrin C. C. Meyer, Melvin Y. Y. Rincon
Summary: This study reports the prevalence of neutralizing antibodies (NAb) against adeno-associated viral (AAV) vectors in Colombian patients with heart failure and healthy individuals. The study found similar prevalence of NAb against different serotypes of AAV vectors in both groups, and showed significantly higher positivity rate for AAV1 and AAV9 antibodies in the heart failure group compared to the control group.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Review
Biotechnology & Applied Microbiology
Dries De Wolf, Kshitiz Singh, Marinee K. Chuah, Thierry Vandendriessche
Summary: Extensive preclinical research has led to the recent regulatory approval of gene therapy products for hemophilia. Roctavian and Hemgenix have shown significant efficacy and safety in clinical trials, resulting in increased levels of clotting factors and reduced bleeding episodes for patients. However, there is variability in patient response and short-term liver inflammation was observed. Longer follow-up studies are needed to determine if lifelong expression of clotting factors can be achieved. Next-generation gene editing technologies offer new prospects for a sustained cure for hemophilia.
HUMAN GENE THERAPY
(2023)
Article
Multidisciplinary Sciences
Jonas Weinmann, Sabrina Weis, Josefine Sippel, Warut Tulalamba, Anca Remes, Jihad El Andari, Anne-Kathrin Herrmann, Quang H. Pham, Christopher Borowski, Susanne Hille, Tanja Schoenberger, Norbert Frey, Martin Lenter, Thierry VandenDriessche, Oliver J. Mueller, Marinee K. Chuah, Thorsten Lamla, Dirk Grimm
NATURE COMMUNICATIONS
(2020)