Article
Oncology
Rui Tian, Jiashuo Liu, Weiwen Fan, Rui Li, Zifeng Cui, Zhuang Jin, Zhaoyue Huang, Hongxian Xie, Lifang Li, Zheying Huang, Zheng Hu, Ping Zhou, Xun Tian
Summary: The development of a gene knockout chain reaction (GKCR) method in this study improves the efficiency of gene editing for the treatment of HPV-related cervical cancer. Through the disruption of the HPV18 E6 and E7 genes using this method, the expression of P53/RB proteins is upregulated and the proliferation and motility of cancer cells are inhibited.
MOLECULAR THERAPY-ONCOLYTICS
(2022)
Article
Fisheries
Jinhai Wang, Baofeng Su, Jacob Al-Armanazi, Allison L. Wise, Mei Shang, Logan Bern, Shangjia Li, De Xing, Andrew Johnson, Wenwen Wang, Darshika U. Hettiarachchi, Michael Coogan, Timothy J. Bruce, Rex A. Dunham
Summary: CRISPR/Cas9-mediated knock-in of the alligator cathelicidin gene into the luteinizing hormone locus in transgenic blue catfish improved disease resistance and showed high integration efficiency. The on-target knock-in efficiency of the double-cut plasmid strategy was higher than that of the linear dsDNA strategy. The As-Cath transgenic blue catfish had significantly higher survival rates during Flavobacterium covae infection compared to wild-type controls.
Article
Biochemistry & Molecular Biology
Kyoka Kuroiwa, Christina Thenault, Fabien Nogue, Laura Perrot, Marianne Mazier, Jean-Luc Gallois
Summary: The susceptibility factor eIF4E2 can be targeted and inactivated in cherry tomato plants using CRISPRCas9-NG. The inactivation of eIF4E2 leads to a narrow resistance spectrum to potyviruses.
Article
Fisheries
De Xing, Baofeng Su, Max Bangs, Shangjia Li, Jinhai Wang, Logan Bern, Rhoda Mae C. Simora, Wenwen Wang, Xiaoli Ma, Michael Coogan, Andrew Johnson, Yi Wang, Zhenkui Qin, Rex Dunham
Summary: This study investigated the use of CRISPR/Cas9 technology for transgenic improvement of channel catfish, showing that knock-in technology can increase the content of n-3 fatty acids. The results indicate that CRISPR/Cas9 technology can more effectively integrate transgenes and enhance the biological effects of foreign genes in fish.
Article
Biotechnology & Applied Microbiology
Rajveer Singh, Shivani Chandel, Arijit Ghosh, Anupam Gautam, Daniel H. Huson, V. Ravichandiran, Dipanjan Ghosh
Summary: In the past two decades, yeast has been widely used as a cost-effective biological tool for producing small molecules and biofuels. The application of CRISPR-Cas techniques in yeast genetic and metabolic engineering has revolutionized the field, particularly in targeted chromosomal integration using synthetic donor constructs. This study presents a highly efficient strategy for targeted chromosomal integration and in-frame expression of foreign genes in the yeast genome using CRISPR-Cas9 and homology-dependent recombination (HDR). The optimized methods achieved a 74% efficiency in integrating small constructs at multiple target sites. The study also demonstrated that as few as 15 base pairs of microhomology are sufficient for targeted knock-in with minimal construct concentration. Whole-genome sequencing confirmed the absence of off-target effects. This protocol provides a comprehensive and streamlined approach for integrating essential genes into the yeast genome for synthetic biology and industrial purposes.
Article
Biochemistry & Molecular Biology
Waheed Shabbir, Nermina Topcagic, Mohammed Aufy, Murat Oz
Summary: This study investigated the role of delta-ENaC subunit in TNF-induced activation of ENaC channel in A549 cells, utilizing CRISPR/Cas 9 knockdown approach and various experimental techniques to analyze the process.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Fisheries
Michael Coogan, Veronica Alston, Baofeng Su, Karim Khalil, Ahmed Elaswad, Mohd Khan, Rhoda M. C. Simora, Andrew Johnson, De Xing, Shangjia Li, Jinhai Wang, Cuiyu Lu, Wenwen Wang, Darshika Hettiarachchi, Tasnuba Hasin, Jeffery Terhune, Ian A. E. Butts, Rex A. Dunham
Summary: This study investigated the effects of CRISPR/Cas9 knockout of the myostatin gene in channel catfish. The results showed that gene-edited channel catfish exhibited high mutation and inheritance rates, as well as improved growth and disease resistance compared to control groups. Therefore, the use of MSTN gene-edited channel catfish is of great importance for commercial farms.
Article
Biochemical Research Methods
Michael Coogan, De Xing, Baofeng Su, Veronica Alston, Andrew Johnson, Mohd Khan, Karim Khalil, Ahmed Elaswad, Shangjia Li, Jinhai Wang, Cuiyu Lu, Wenwen Wang, Darshika Hettiarachchi, Mei Shang, Tasnuba Hasin, Zhenkui Qin, Roger Cone, Ian A. E. Butts, Rex A. Dunham
Summary: By using genome engineering, three transgenes (masu salmon elongase gene, common carp beta-actin promoter, and melanocortin-4 receptor gene) were inserted into the channel catfish genome, resulting in improved growth and increased omega-3 fatty acid content. This is the first successful example of applying genome engineering to improve commercially important aquaculture species. Transgenic channel catfish with Elovl2 gene show potential for application in commercial farms.
TRANSGENIC RESEARCH
(2023)
Article
Chemistry, Multidisciplinary
Ruosen Xie, Xiuxiu Wang, Yuyuan Wang, Mingzhou Ye, Yi Zhao, Brian S. Yandell, Shaoqin Gong
Summary: This study reports the development of a facilely fabricated pH-responsive polymer nanoparticle for safe and efficient delivery of CRISPR-associated protein 9 (Cas9) for gene editing. These nanoparticles possess many desirable properties, making them potential candidates for clinical applications in genome editing.
ADVANCED MATERIALS
(2022)
Article
Biochemistry & Molecular Biology
Md. Lutfur Rahman, Toshinori Hyodo, Muhammad Nazmul Hasan, Yuko Mihara, Sivasundaram Karnan, Akinobu Ota, Shinobu Tsuzuki, Yoshitaka Hosokawa, Hiroyuki Konishi
Summary: This study developed a sensitive assay, named the CD55 correction assay, to quantify the efficiency of targeted knock-in. The assay allows high-throughput, quantitative detection of targeted gene correction events occurring in an endogenous human gene using flow cytometry.
MOLECULAR BIOLOGY REPORTS
(2022)
Article
Biochemistry & Molecular Biology
Shuai Wang, Chen Tao, Huilin Mao, Linghui Hou, Yao Wang, Tao Qi, Yuan Yang, Sang-Ging Ong, Shijun Hu, Renjie Chai, Yongming Wang
Summary: Due to different nucleotide preferences at target sites, no single Cas9 is capable of editing all sequences. Thus, there is a need to establish a Cas9 repertoire covering all sequences for efficient genome editing. In this study, we identified new SaCas9 orthologs with a simple NNGG PAM requirement by comparing the PAM-interacting domains in 16 SaCas9 orthologs. We also discovered three amino acids that determine the NNGG PAM requirement of SaCas9. Furthermore, we engineered high-fidelity versions of Cas9s, Sha2Cas9 and SpeCas9, which displayed high activities and distinct nucleotide preferences. Our findings offer a new perspective for expanding the Cas9 repertoire and identifying SaCas9 orthologs with NNGG PAM requirements.
Article
Materials Science, Multidisciplinary
Yue Wang, Xiujue Zheng, Jun Liu, Qixian Chen, Yan Zhao
Summary: The emergence of CRISPR-based genome editing systems has revolutionized the field of molecular biology and holds great promise for the treatment of various diseases. However, the lack of suitable delivery vehicles for effective intracellular delivery of CRISPR components limits its practical applications. In this study, a nanometer-scaled vesicle platform (CRISPRsome) was developed to efficiently deliver the essential components of the CRISPR system, resulting in significant gene knockout efficacy in cancer cells. This study highlights the potential of CRISPRsome in facilitating the use of CRISPR technology in practical applications.
ACS MATERIALS LETTERS
(2022)
Article
Biochemical Research Methods
Saori Mizuno-Iijima, Shinya Ayabe, Kanako Kato, Shogo Matoba, Yoshihisa Ikeda, Tra Thi Huong Dinh, Hoai Thu Le, Hayate Suzuki, Kenichi Nakashima, Yoshikazu Hasegawa, Yuko Hamada, Yoko Tanimoto, Yoko Daitoku, Natsumi Iki, Miyuki Ishida, Elzeftawy Abdelaziz Elsayed Ibrahim, Toshiaki Nakashiba, Michito Hamada, Kazuya Murata, Yoshihiro Miwa, Miki Okada-Iwabu, Masato Iwabu, Ken-ichi Yagami, Atsuo Ogura, Yuichi Obata, Satoru Takahashi, Seiya Mizuno, Atsushi Yoshiki, Fumihiro Sugiyama
Summary: Genetically modified mouse models are crucial for studying gene function and human diseases. By incorporating the nuclear localization property of Cdt1 protein into the CRISPR-Cas system, Cas9-mC was able to efficiently produce mice carrying targeted mutations, including bi-allelic full deletions and fragment knock-ins.
Article
Endocrinology & Metabolism
Divya Ramachandran, Kusum Sharma, Vishal Saxena, Niepukolie Nipu, Dinusha C. Rajapaksha, Jan A. Mennigen
Summary: The nonapeptide vasotocin/vasopressin is highly conserved and plays a role as neuromodulator and signaling molecule in vertebrates. Pharmacological studies in several teleost fishes suggest that vasotocin has sex and species-specific reproductive roles. However, comprehensive studies investigating the behavioral and physiological consequences of genetic ablation of vasotocin in a genetically tractable fish model, such as the zebrafish, are currently lacking.
FRONTIERS IN ENDOCRINOLOGY
(2023)
News Item
Multidisciplinary Sciences
Smriti Mallapaty
Summary: Concerns about excessive interference are overshadowing the proposal for protecting children whose genomes were edited.
Article
Fisheries
Muyassar Abualreesh, Jaelen N. Myers, Jeremy Gurbatow, Andrew Johnson, De Xing, Jinhai Wang, Shangjia Li, Michael Coogan, Khoi Vo, Nour El Husseini, Rex A. Dunham, Ian A. E. Butts
Summary: This study demonstrated that combining specific antioxidants and AFPs can improve spermatogonia cryopreservation for a fish species, providing recommendations for future cryopreservation protocols. While individual antioxidant or AFP treatments did not show improvements, specific combinations resulted in higher cell production and viability.
Article
Genetics & Heredity
Xiao Xiong, Paul B. Samollow, Wenqi Cao, Richard Metz, Chao Zhang, Ana C. Leandro, John L. VandeBerg, Xu Wang
Summary: The study characterized the genetic profile of laboratory strains of the gray short-tailed opossum, identifying a large number of high-quality SNPs and confirming the successful establishment of inbred strains with different degrees of inbreeding. Population structure analysis reflected the breeding history of these strains and indicated their suitability for various experimental protocols and genetic studies.
G3-GENES GENOMES GENETICS
(2022)
Article
Biology
Haolong Wang, Timothy J. Bruce, Baofeng Su, Shangjia Li, Rex A. Dunham, Xu Wang
Summary: Hematological assays and transcriptome analysis were performed in the hybrid catfish to investigate the mechanisms of environment-dependent heterosis. The study found that the channel catfish showed lower innate immunity activity, stress, and blood glucose/lactate levels, which are associated with its superior growth performance. Genes involved in fatty acid metabolism/transport pathways were significantly upregulated in channel catfish. The research also revealed that hybrids have increased blood glucose levels due to transgressive expression. This study provides insights into the molecular mechanisms of heterosis and has implications for genetic enhancement through hybrid breeding.
Article
Fisheries
Nermeen Y. Abass, Rhoda Mae C. Simora, Jinhai Wang, Shangjia Li, De Xing, Michael Coogan, Andrew Johnson, David Creamer, Xu Wang, Rex A. Dunham
Summary: This study demonstrates that genetic enhancement of the channel catfish's resistance to Edwardsiella ictaluri can be achieved through cecropin transgenesis. The transgenic fish exhibited enhanced disease resistance and improved survival time when challenged with the bacteria. The growth rates of transgenic and non-transgenic fish were not different, indicating that the transgene did not affect the overall growth of the fish.
FISH & SHELLFISH IMMUNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
De Xing, Baofeng Su, Shangjia Li, Max Bangs, David Creamer, Michael Coogan, Jinhai Wang, Rhoda Simora, Xiaoli Ma, Darshika Hettiarachchi, Veronica Alston, Wenwen Wang, Andrew Johnson, Cuiyu Lu, Tasnuba Hasin, Zhenkui Qin, Rex Dunham
Summary: Omega-3 polyunsaturated fatty acids, particularly EPA and DHA, play a crucial role in human health. In this study, a transgenic approach was used to enhance the EPA and DHA content in channel catfish by inserting the elovl2 gene from masu salmon. The results showed promising improvements, indicating the potential for enhancing n-3 PUFAs in channel catfish.
MARINE BIOTECHNOLOGY
(2022)
Article
Genetics & Heredity
Rebecca L. Nance, Sara J. Cooper, Dmytro Starenki, Xu Wang, Brad Matz, Stephanie Lindley, Annette N. Smith, Ashley A. Smith, Noelle Bergman, Maninder Sandey, Jey Koehler, Payal Agarwal, Bruce F. Smith
Summary: Despite efforts to improve patient survival in osteosarcoma (OSA), an aggressive bone tumor, the complexity of its genetics, drug resistance, and heterogeneity pose challenges for single-target treatments. Precision medicine aims to address the intra- and inter-tumoral heterogeneity to enhance patient outcome. By characterizing differentially expressed genes (DEGs) unique to the tumor, appropriate therapies and novel treatments can be developed. Canine OSA, which shares similarities with the human disease, serves as a useful model for studying DEGs at the individual level.
Article
Biology
A. K. Shakur Ahammad, Neaz A. Hasan, Abul Bashar, Mohammad Mahfujul Haque, Muyassar H. Abualreesh, Md Mehefuzul Islam, Biraj Kumar Datta, Md Fazla Rabbi, Mohd Golam Quader Khan, Md Samsul Alam
Summary: The objective of this study was to evaluate the growth performance and genetic variation in diallel crosses of Ariza labeo originating from three geographically separated rivers in Bangladesh. The results showed that the G4K female A male group had higher fertilization, hatching, and survival rates compared to other groups. This group also exhibited better length and weight gains, SGR, and heterosis. Additionally, they had a higher number of hyperplastic muscle fibers and genetic diversity. Therefore, the G4K female A male group is considered for continuing the L. ariza stock improvement program.
Article
Fisheries
Michael Coogan, Veronica Alston, Baofeng Su, Karim Khalil, Ahmed Elaswad, Mohd Khan, Rhoda M. C. Simora, Andrew Johnson, De Xing, Shangjia Li, Jinhai Wang, Cuiyu Lu, Wenwen Wang, Darshika Hettiarachchi, Tasnuba Hasin, Jeffery Terhune, Ian A. E. Butts, Rex A. Dunham
Summary: This study investigated the effects of CRISPR/Cas9 knockout of the myostatin gene in channel catfish. The results showed that gene-edited channel catfish exhibited high mutation and inheritance rates, as well as improved growth and disease resistance compared to control groups. Therefore, the use of MSTN gene-edited channel catfish is of great importance for commercial farms.
Article
Fisheries
De Xing, Baofeng Su, Max Bangs, Shangjia Li, Jinhai Wang, Logan Bern, Rhoda Mae C. Simora, Wenwen Wang, Xiaoli Ma, Michael Coogan, Andrew Johnson, Yi Wang, Zhenkui Qin, Rex Dunham
Summary: This study investigated the use of CRISPR/Cas9 technology for transgenic improvement of channel catfish, showing that knock-in technology can increase the content of n-3 fatty acids. The results indicate that CRISPR/Cas9 technology can more effectively integrate transgenes and enhance the biological effects of foreign genes in fish.
Article
Fisheries
Andrew Johnson, Katherine Zipfel, Eric Hallerman, Wade Massure, Peter Euclide, Amy Welsh
Summary: The objective of this study was to create a faster and more accurate genetic assignment tool for Walleye broodstock in the Eastern Highlands region and to measure the genetic diversity of four Walleye populations using next-generation sequencing. The results showed that the four populations were genetically distinct from each other and each population had varying degrees of genetic differentiation from its source population. The study also identified a pure native Eastern Highlands strain that can be used for future augmentation and restoration efforts.
TRANSACTIONS OF THE AMERICAN FISHERIES SOCIETY
(2023)
Article
Fisheries
Darshika Udari Hettiarachchi, Veronica N. Alston, Jeremy Gurbatow, Hana N. Hess, Baofeng Su, Andrew Johnson, Michael P. Coogan, Jinhai Wang, De Xing, Shangjia Li, David Creamer, Vahid Zadmajid, Ian A. E. Butts, Rex A. Dunham
Summary: Xenogenesis is an innovative tool for hybrid catfish seed production accomplished by transplanting undifferentiated germ cells from a donor fish into a sterile recipient. This study aimed to evaluate seasonal variations in germ cell counts and serum sex steroid hormonal profiles in mature Blue Catfish. The results showed that the counts of live germ cells and levels of serum sex steroid hormones varied seasonally.
NORTH AMERICAN JOURNAL OF AQUACULTURE
(2023)
Article
Fisheries
Mary Akers, Henry Quinlan, Andrew Johnson, Edward Baker, Amy Welsh
Summary: The objective of this study was to compare paternal representation and genetic diversity between the two stocking strategies. Parentage analysis based on genetic data determined that none of the family groups in the hatchery had equal paternal representation, while wild-produced offspring had equal paternal representation. Despite the larger number of breeders contributing to the wild-caught larvae, there was no significant difference in genetic diversity between the wild-caught larvae and representative hatchery-produced offspring.
Article
Biology
Yihang Zhou, Yi Wang, Xiao Xiong, Arthur G. Appel, Chao Zhang, Xu Wang
Summary: Telomeres are composed of highly conserved simple tandem telomeric repeat motifs, which vary among different species. A fast Telomeric Repeats Identification Pipeline was developed and its accuracy was demonstrated in multiple species. The study reveals diverse telomeric repeat motifs in different insect species and rapid evolution of telomeres.
LIFE SCIENCE ALLIANCE
(2022)
