Review
Biochemistry & Molecular Biology
Shuai Ding, Jinfeng Liu, Xin Han, Mengfan Tang
Summary: The CRISPR/Cas9 system, a highly efficient and accurate gene editing tool, has been widely applied in cancer research. Its utilization in CAR-T-cell therapy, tumor model establishment, and gene and drug target screening has propelled the investigation of cancer molecular mechanisms and the advancement of precision medicine. However, the therapeutic potential of genome editing remains underexplored and may pose risks of additional genetic mutations.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Review
Chemistry, Multidisciplinary
Peng Yang, Athena Yue-Tung Lee, Jingjing Xue, Shih-Jie Chou, Calvin Lee, Patrick Tseng, Tiffany X. Zhang, Yazhen Zhu, Junseok Lee, Shih-Hwa Chiou, Hsian-Rong Tseng
Summary: The CRISPR/Cas9 genome editing system has revolutionized the field of therapeutic applications for genetic diseases and cancers. Efficient delivery of the system in vivo remains a significant challenge. Non-viral nano-vectors offer advantages over viral vectors and can be used to deliver CRISPR/Cas9 cargoes for targeted gene editing. The article highlights recent advances in nano-vector delivery for treating cancers and genetic diseases and proposes strategies for future development.
Article
Biochemistry & Molecular Biology
Jiyeon Kweon, An-Hee Jang, Eunji Kwon, Ungi Kim, Ha Rim Shin, Jieun See, Gayoung Jang, Chaeyeon Lee, Taeyoung Koo, Seokjoong Kim, Yongsub Kim
Summary: This article introduces the use of various CRISPR-Cas9 orthologs in genome engineering. One of the smallest Cas9 orthologs, cjCas9, derived from Campylobacter jejuni, has been developed into base editors capable of inducing endogenous base substitutions with high efficiency and specificity. The engineered cjCas9 and gRNA scaffolds can further improve the base editing efficiency of cjABE8e. The findings in this study expand the potential of using base editors for therapeutic approaches.
EXPERIMENTAL AND MOLECULAR MEDICINE
(2023)
Article
Multidisciplinary Sciences
Yeh-Hsing Lao, Robin Ji, Joyce K. Zhou, Kathy J. Snow, Nancy Kwon, Ethan Saville, Siyu He, Shradha Chauhan, Chun-Wei Chi, Malika S. Datta, Hairong Zhang, Chai Hoon Quek, S. Sarah Cai, Mingqiang Li, Yaned Gaitan, Lawrence Bechtel, Shih-Ying Wu, Cathleen M. Lutz, Raju Tomer, Stephen A. Murray, Alejandro Chavez, Elisa E. Konofagou, Kam W. Leong
Summary: This study demonstrates a safe and effective gene editing technique by using focused ultrasound to transiently open the blood-brain barrier for the transport of CRISPR/Cas9 machinery to the brain.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2023)
Article
Biochemical Research Methods
Wenyi Wu, Yanhui Yang, Fei Yao, Lijun Dong, Xiaobo Xia, Shaochong Zhang, Hetian Lei
Summary: This method delivers CRISPR-Cas9 into vascular endothelial cells using AAVs, with SpCas9 expression driven by a specific endothelial promoter to restrict the foreign enzyme in these cells. By editing VEGFR2 in retinal vascular endothelial cells in a mouse model, this simplified protocol can potentially be expanded to other research on editing endothelial genome in vivo.
Article
Biochemistry & Molecular Biology
Xiaopeng Song, Yaxiong Cui, Yanxiao Wang, Yizhe Zhang, Qi He, Zhenyang Yu, Chengfang Xu, Huimin Ning, Yuying Han, Yunting Cai, Xuan Cheng, Jian Wang, Yan Teng, Xiao Yang, Jun Wang
Summary: In this study, the AAV-BR1-CRISPR system was used to specifically knock out target genes in brain endothelial cells of adult mice, demonstrating its effectiveness in rapidly identifying endothelial genes that regulate blood-brain barrier integrity in vivo.
INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES
(2022)
Article
Biochemistry & Molecular Biology
Hong You, Johannes U. Mayer, Rebecca L. Johnston, Haran Sivakumaran, Shiwanthi Ranasinghe, Vanessa Rivera, Olga Kondrashova, Lambros T. Koufariotis, Xiaofeng Du, Patrick Driguez, Juliet D. French, Nicola Waddell, Mary G. Duke, Wannaporn Ittiprasert, Victoria H. Mann, Paul J. Brindley, Malcolm K. Jones, Donald P. McManus
Summary: CRISPR/Cas9-mediated genome editing has shown great potential for genetic modification of helminth parasites, as demonstrated by successful gene knock-in in Schistosoma mansoni eggs. This study confirmed the utility of CRISPR/Cas9 for functional genomics in schistosomes through combining CRISPR/Cas9 with single-stranded oligodeoxynucleotides (ssODNs).
Article
Agricultural Engineering
Xuemei Dai, Xianfeng Yang, Chun Wang, Yueting Fan, Shichao Xin, Yuwei Hua, Kejian Wang, Huasun Huang
Summary: In this study, CRISPR/Cas9 system was established in Hevea brasiliensis using five endogenous U6 promoters to drive sgRNA transcription. Mutation patterns were characterized in protoplasts targeting flowering time related genes, and insertion, deletion, and base substitution mutations were detected. A stable transformation editing vector targeting the phytoene desaturase gene was constructed and induced albino phenotype in rubber tree callus. This study demonstrates successful establishment of genome editing in Hevea brasiliensis using CRISPR/Cas9 plasmid system.
INDUSTRIAL CROPS AND PRODUCTS
(2021)
Article
Biology
Jian-Ping Zhang, Zhi-Xue Yang, Feng Zhang, Ya-Wen Fu, Xin-Yue Dai, Wei Wen, Beldon Zhang, Hannah Choi, Wanqiu Chen, Meredith Brown, David Baylink, Lei Zhang, Hongyu Qiu, Charles Wang, Tao Cheng, Xiao-Bing Zhang
Summary: Research shows that HDAC inhibitors can increase the efficiency of genome editing, especially when dealing with silent genes. HDR efficiency improvement is more significant at closed loci.
SCIENCE CHINA-LIFE SCIENCES
(2021)
Article
Multidisciplinary Sciences
Joonbum Lee, Dong-Hwan Kim, Madeline C. Karolak, Sangsu Shin, Kichoon Lee
Summary: In this study, we successfully generated genome-edited chicken and duck lines without the conventional PGC-mediated procedures by injecting an adenovirus containing the CRISPR-Cas9 system into avian blastoderms.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2022)
Article
Plant Sciences
Xinrong Li, Xin Zuo, Mingming Li, Xu Yang, Jingyu Zhi, Hongzheng Sun, Caixia Xie, Zhongyi Zhang, Fengqing Wang
Summary: This study cloned a PDS gene from Rehmannia glutinosa, achieved gene editing using CRISPR/Cas9 system to generate albino plants, providing a foundation for further research and improvement of R. glutinosa varieties.
PLANT CELL REPORTS
(2021)
Article
Biochemical Research Methods
Xuemi Hao, Tingzhen Mu, Moustafa Mohamed Sharshar, Yunpu Jia, Wei Zhong, Zheng Chen, Qifeng Wen, Maohua Yang, Caixia Wang, Jianmin Xing
Summary: In this study, a CRISPR/AsCas12a-mediated system was developed in T. versutus for efficient genome editing. Compared to the CRISPR/Cas9 system, the CRISPR/AsCas12a system exhibited enhanced editing efficiency and allowed multiplex gene editing and large-fragment DNA knockout. By using this system, five key genes of the elemental sulfur oxidation pathway were successfully knocked out, resulting in improved understanding of sulfur metabolism in Thioalkalivibrio spp.
ACS SYNTHETIC BIOLOGY
(2023)
Article
Biochemical Research Methods
Xuemi Hao, Tingzhen Mu, Moustafa Mohamed Sharshar, Yunpu Jia, Wei Zhong, Zheng Chen, Qifeng Wen, Maohua Yang, Caixia Wang, Jianmin Xing
Summary: In this study, a CRISPR/AsCas12a-mediated system was developed for efficient and implementable genome editing workflow in Thioalkalivibrio versutus. Compared to the CRISPR/Cas9 system, the CRISPR/AsCas12a system exhibited enhanced editing efficiency and allowed multiplex gene editing and large-fragment DNA knockout using one promoter. Knocking out five key genes of the elemental sulfur oxidation pathway disrupted the ability of T. versutus to metabolize elemental sulfur, leading to an increase in elemental sulfur generation and a reduction in sulfate production. This genome engineering strategy significantly improved our understanding of sulfur metabolism in Thioalkalivibrio spp.
ACS SYNTHETIC BIOLOGY
(2023)
Article
Oncology
Imran Ansari, Animesh Chaturvedi, Deepak Chitkara, Saurabh Singh
Summary: The understanding of the relationship between epigenetic alterations and gene expression have opened up new therapeutic pathways for treating various diseases, including cancer. CRISPR/Cas and other genome editing technologies have shown potential in epigenome editing. This review outlines the epigenetic therapeutic approach using CRISPR/Cas for cancer therapy.
SEMINARS IN CANCER BIOLOGY
(2022)
Article
Plant Sciences
Pawan Mainkar, Tushar Kashinath Manape, Viswanathan Satheesh, Sivalingam Anandhan
Summary: This study reports the successful establishment of a CRISPR/Cas9-mediated genome editing protocol in onions, using the AcPDS gene as an example. Through transformation and cultivation of calli, different phenotypes were observed in regenerated shoots, with some exhibiting albino or chimeric characteristics. PCR and deep amplicon sequencing confirmed the successful editing of the AcPDS gene.
FRONTIERS IN PLANT SCIENCE
(2023)
Article
Biochemical Research Methods
Lijie Zhang, Rongyao Dong, Robert J. Zawadzki, Pengfei Zhang
Summary: Optoretinogram, utilizing OCT to measure retinal functions, is a potentially useful tool for quantifying retinal health alterations. This study improved accuracy by collecting volumetric data and developing a 3D registration approach, revealing novel functional signals and enabling monitoring of local and global functional changes in rodent eyes.
JOURNAL OF BIOPHOTONICS
(2022)
Review
Optics
Danuta M. Sampson, Adam M. Dubis, Fred K. Chen, Robert J. Zawadzki, David D. Sampson
Summary: The visualization and assessment of retinal microvasculature are crucial in the study and treatment of eye and systemic diseases. However, the current methods and analysis of OCTA imaging lack consistency, hindering progress. This paper proposes steps to standardize OCTA imaging, including imaging protocols, data analysis methods, metrics, reporting, and clinical practice, to address these limitations.
LIGHT-SCIENCE & APPLICATIONS
(2022)
Article
Medicine, Research & Experimental
Sook Hyun Chung, Tzu-Ni Sin, Brian Dang, Taylor Ngo, Therlinder Lo, Daniella Lent-Schochet, Ratheesh K. Meleppat, Robert J. Zawadzki, Glenn Yiu
Summary: This study compares the effectiveness of using single versus paired gRNAs in the CRISPR-Cas9 system to target the Vegfa gene. Paired gRNAs increased gene ablation rates in human cells but did not enhance VEGF suppression in mouse eyes. Using two gRNAs for genome editing may increase the risk of off-target effects.
MOLECULAR THERAPY-NUCLEIC ACIDS
(2022)
Article
Ophthalmology
Ratheesh K. Meleppat, Christopher R. Fortenbach, Yifan Jian, Esteban Soto Martinez, Karen Wagner, Bobeck S. Modjtahedi, Monica J. Motta, Deepa L. Ramamurthy, Ivan R. Schwab, Robert J. Zawadzki
Summary: In vivo evaluation of the structural morphology and vascular plexuses of the neurosensory retina and choroid across vertebrate species was performed using SS-OCT and SS-OCTA imaging. The results showed that the retinal morphology and vascular plexuses varied among different species. These methods can be used to evaluate retinal diseases in various species.
TRANSLATIONAL VISION SCIENCE & TECHNOLOGY
(2022)
Article
Optics
Kari V. Vienola, Denise Valente, Robert J. Zawadzki, Ravi S. Jonnal
Summary: Optoretinography (ORG) is a noninvasive and objective tool for testing neural function in the retina. It has the potential to transform ophthalmic care and clinical trials of therapeutics for visual function. The article presents an alternative approach that monitors the velocity of retinal features instead of tracking their positions, reducing the complexity and cost of position-based methods.
Article
Optics
Soohyun Lee, Stacey S. Choi, Ratheesh K. Meleppat, Robert J. Zawadzki, Nathan Doble
Summary: This article describes a high-speed, adaptive optics partially confocal multi-spot ophthalmoscope (AO-pcMSO) that utilizes a digital micromirror device (DMD) in the illumination channel and a fast 2D CMOS camera. The camera is synchronized with the DMD to project multiple AO-corrected spots onto the human retina simultaneously. Spatial filtering on each raw retinal image acts as an array virtual pinholes. The parallel projection scheme achieves a frame acquisition rate of 250 fps and improves contrast by 2-3 fold compared to a standard flood illumination architecture. Partially confocal images of the human retina reveal cone and rod photoreceptors at various retinal eccentricities.
Article
Biochemical Research Methods
Pengfei Zhang, Daniel J. Wahl, Jacopo Mocci, Eric B. Miller, Stefano Bonora, Marinko Sarunic, Robert J. Zawadzki
Summary: Optical coherence tomography (OCT) and scanning laser ophthalmoscopy (SLO), invented in the 1980s, have greatly advanced in vivo retinal diagnostics in ophthalmology clinics and vision science research. Adaptive optics (AO) technology improves the resolution and sensitivity of SLO and OCT systems by correcting ocular aberrations. Integrating OCT into an existing mouse retinal AO-SLO system allows for multi-modal AO-enhanced imaging of the living mouse eye with high acquisition speed and resolution.
BIOMEDICAL OPTICS EXPRESS
(2023)
Meeting Abstract
Ophthalmology
Ewelina Pijewska, Kari V. Vienola, Michal Meina, Pengfei Zhang, Ratheesh K. Meleppat, Maciej Szkulmowski, Ravi S. Jonnal, Robert J. Zawadzki
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
(2022)
Meeting Abstract
Ophthalmology
Christopher Loh, Pengfei Zhang, Ratheesh K. Meleppat, Oscar Ramos-Soto, Sandra E. Balderas-Mata, Robert J. Zawadzki, Susanna S. Park
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
(2022)
Meeting Abstract
Ophthalmology
Ratheesh K. Meleppat, Kaitryn Ronning, Sarah J. Karlen, J. KarlenMarie E. Burns, Robert J. Zawadzki
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
(2022)
Meeting Abstract
Ophthalmology
Jessicca Cho, Pengfei Zhang, Sarah J. Karlen, Nicholas Marsh-Armstrong, Anna La Torre, Robert J. Zawadzki
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
(2022)