Article
Multidisciplinary Sciences
Joshua R. Elmore, Gara N. Dexter, Henri Baldino, Jay D. Huenemann, Ryan Francis, V. George L. Peabody, Jessica Martinez-Baird, Lauren A. Riley, Tuesday Simmons, Devin Coleman-Derr, Adam M. Guss, Robert G. Egbert
Summary: SAGE is a highly efficient and extensible genome engineering technology that allows selection marker-free, site-specific genome integration of multiple DNA constructs in bacteria. It shows superior efficiency compared to replicating plasmids and overcomes host range limitations. SAGE has the potential to expand high-throughput genetics and synthetic biology in industrial and environmental bacteria.
Article
Cell Biology
Yingqi Lin, Caijuan Li, Wei Wang, Jiawei Li, Chunhui Huang, Xiao Zheng, Zhaoming Liu, Xichen Song, Yizhi Chen, Jiale Gao, Jianhao Wu, Jiaxi Wu, Zhuchi Tu, Liangxue Lai, Xiao-Jiang Li, Shihua Li, Sen Yan
Summary: Animal models are crucial for understanding human diseases and developing drugs. Pigs are extensively used in studying various systems and have become an attractive model for neurodegenerative diseases. This study demonstrates that intravenous injection of AAV9-GFP leads to widespread transgene expression in various organs, including the brain, without causing inflammation. These findings suggest that intravenous administration of AAV9 can be used to establish large animal models for neurodegenerative diseases and to develop treatments.
FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY
(2023)
Article
Genetics & Heredity
Nawaphat Malaiwong, Montserrat Porta-de-la-Riva, Michael Krieg
Summary: The stable incorporation of transgenes and recombinant DNA material into the host genome is a challenge in bioengineering. A library of 300 different loci in Caenorhabditis elegans provides safe harbors for transgene integration using CRISPR/Cas9. This method allows easy tracking of transgene integration and can be used without affecting body morphology or behavior.
G3-GENES GENOMES GENETICS
(2023)
Article
Genetics & Heredity
Xiang Ma, Weijun Zeng, Lei Wang, Rui Cheng, Zeying Zhao, Caiyun Huang, Zhongxin Sun, Peipei Tao, Tao Wang, Jufang Zhang, Lu Liu, Xing Duan, Dong Niu
Summary: Transgenic technology is widely used in biomedical and agricultural fields, but random integration can cause uncertain consequences. This study aimed to identify the best safe harbor (SH) locus for efficient porcine transgenesis. The results showed that COL1A1 and ROSA26 were the most favorable SH loci, with COL1A1 being more competitive. This research will greatly facilitate porcine genome engineering and transgenic pig production.
FUNCTIONAL & INTEGRATIVE GENOMICS
(2022)
Article
Agriculture, Multidisciplinary
Guang-Ming Xiang, Xiu-ling Zhang, Chang-jiang Xu, Zi-yao Fan, Kui Xu, Nan Wang, Yue Wang, Jing-jing Che, Song-song Xu, Yu-lian Mu, Kui Li, Zhi-guo Liu
Summary: A study showed that efficient transgene knock-in using the CRISPR/Cas9 system in the COL1A1 gene did not affect mRNA expression or global gene expression, and had no adverse effects on cell proliferation. This indicates that the COL1A1 locus can serve as a safe harbor for the insertion of foreign genes.
JOURNAL OF INTEGRATIVE AGRICULTURE
(2023)
Article
Genetics & Heredity
Sonia El Mouridi, Faisal Alkhaldi, Christian Frokjaer-Jensen
Summary: The MosTI technique enables efficient and reproducible transgenesis in Caenorhabditis elegans, facilitating targeted insertion of single-copy transgenes and integration of extrachromosomal arrays. The technology allows for easy conversion between different selection markers and provides a collection of universal targeting vectors with commonly used promoters and fluorophores. Transgenes are expressed consistently in somatic and germ cells, and chromosomal integration is mediated by CRISPR/Cas9.
G3-GENES GENOMES GENETICS
(2022)
Article
Multidisciplinary Sciences
Adriana Adolfi, Amy Lynd, Gareth J. Lycett, Anthony A. James
Summary: The phi C31 system is a crucial genetic tool for the transgenic manipulation of malaria mosquitoes and other insect vectors, enabling precise modification of the Anopheles genome to achieve genetic control of malaria.
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
(2021)
Article
Biotechnology & Applied Microbiology
Sana Pourtabatabaei, Samaneh Ghanbari, Narges Damavandi, Elham Bayat, Mozhgan Raigani, Sirous Zeinali, Fatemeh Davami
Summary: Chinese hamster ovary (CHO) cells are widely used for manufacturing therapeutic proteins, and in this study, stable cell lines were successfully generated with targeted integration of reporter genes using the CRISPR/Cas9 platform. The study showed that higher efficiency and expression homogeneity were observed in the pseudo attP site associated with chromosome 6 compared to the pseudo attP site from chromosome 3.
JOURNAL OF BIOTECHNOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Maria H. T. Nguyen, Zoe Netter, Angus Angermeyer, Kimberley D. Seed
Summary: Bacteria can acquire mobile genetic elements to combat viral infection. Satellite viruses restrict viral replication for the benefit of their host bacterium. In the arms race against satellite viruses, certain viruses can evade restriction through unknown mechanisms. This study identifies a viral protein that counteracts satellite viruses by exploiting a specific enzyme within the viruses.
NUCLEIC ACIDS RESEARCH
(2022)
Article
Biochemistry & Molecular Biology
Christian Sudfeld, Ana Pozo-Rodriguez, Sara A. Manjavacas Diez, Rene H. Wijffels, Maria J. Barbosa, Sarah D'Adamo
Summary: This study reports a novel gene expression system discovered in the oleaginous microalga Nannochloropsis, utilizing the highly efficient transcriptional activity of RNA polymerase I and an internal ribosome entry site for translation. By identifying the nucleolus as a genomic safe harbor for Pol I transcription and constructing transformant strains, consistently strong transgene expression was achieved, providing a powerful tool for genetic and metabolic engineering of microalgae and potential significant contributions to microalgal research.
Article
Economics
Alessandro De Chiara, Ester Manna
Summary: This study examines whether the combined adoption of ex-ante regulation and ex-post liability leads to a higher level of welfare when firms invest in the development of innovative products with potential negative social impacts. The presence of firm-regulator corruption is taken into account, and two alternative regulatory regimes, lenient authorization and strict authorization, are compared. The findings indicate that corruption favors strict authorization and reinforces the argument for making firms immune from ex-post liability to encourage ex-ante investment. However, when lenient authorization is adopted, firms should not be insulated from liability, implying that liability should be more severe in situations with less common corruption.
Review
Biotechnology & Applied Microbiology
Vishnu Hosur, Benjamin E. Low, Michael V. Wiles
Summary: The inability to efficiently and precisely insert large DNA constructs into the genome is a major challenge in genomic engineering. Existing methods lack precision or have limited capacity. However, a new toolkit combining RNA-guided Cas9 and site-specific integrase Bxb1 has been developed, enabling the integration of DNA constructs ranging from 5 to 43 kb in mouse zygotes and human cells.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Matthew G. Durrant, Alison Fanton, Josh Tycko, Michaela Hinks, Sita S. Chandrasekaran, Nicholas T. Perry, Julia Schaepe, Peter P. Du, Peter Lotfy, Michael C. Bassik, Lacramioara Bintu, Ami S. Bhatt, Patrick D. Hsu
Summary: Large serine recombinases (LSRs) are DNA integrases that facilitate the site-specific integration of mobile genetic elements into bacterial genomes. In this study, a computational approach was developed to identify thousands of LSRs and their DNA attachment sites, expanding known LSR diversity and enabling more efficient DNA integration. This research provides new tools for inserting large sequences into the human genome.
NATURE BIOTECHNOLOGY
(2023)
Article
Multidisciplinary Sciences
John J. Kelly, Moe Saee-Marand, Nivin N. Nystrom, Melissa M. Evans, Yuanxin Chen, Francisco M. Martinez, Amanda M. Hamilton, John A. Ronald
Summary: The study utilized HITI CRISPR-Cas9 minicircle donors for precise targeted knock-in of reporter genes at safe harbor loci. Results showed higher knock-in efficiency and functional reporter gene activity with HITI vectors, enabling multi-modal longitudinal in vivo imaging of cells. This work demonstrates the first CRISPR-Cas9 HITI system for large DNA donor constructs at a safe harbor locus.
Article
Multidisciplinary Sciences
Chee-Hong Takahiro Yew, Narmatha Gurumoorthy, Fazlina Nordin, Gee Jun Tye, Wan Safwani Wan Kamarul Zaman, Jun Jie Tan, Min Hwei Ng
Summary: HIV-1-derived lentiviral vector is an efficient transporter for delivering desired genetic materials into targeted cells. Integrase-deficient lentiviral vector, with reduced integration rate, offers better safety and wider clinical applications.
Article
Multidisciplinary Sciences
Stefan Wagner, Rosemary Thresher, Ross Bland, Goetz Laible
SCIENTIFIC REPORTS
(2015)
Review
Biochemical Research Methods
Mark Tizard, Eric Hallerman, Scott Fahrenkrug, Martina Newell-McGloughlin, John Gibson, Frans de Loos, Stefan Wagner, Gotz Laible, Jae Yong Han, Michael D'Occhio, Lisa Kelly, John Lowenthal, Kari Gobius, Primal Silva, Caitlin Cooper, Tim Doran
TRANSGENIC RESEARCH
(2016)
Article
Multidisciplinary Sciences
Yanzhen Bi, Zaidong Hua, Ximei Liu, Wenjun Hua, Hongyan Ren, Hongwei Xiao, Liping Zhang, Li Li, Zhirui Wang, Gotz Laible, Yan Wang, Faming Dong, Xinmin Zheng
SCIENTIFIC REPORTS
(2016)
Article
Multidisciplinary Sciences
Jingwei Wei, Stefan Wagner, Paul Maclean, Brigid Brophy, Sally Cole, Grant Smolenski, Dan F. Carlson, Scott C. Fahrenkrug, David N. Wells, Gotz Laible
SCIENTIFIC REPORTS
(2018)
Article
Multidisciplinary Sciences
Gotz Laible, Grant Smolenski, Thomas Wheeler, Brigid Brophy
SCIENTIFIC REPORTS
(2016)
Article
Multidisciplinary Sciences
Medhat A. Al-Ghobashy, Aliaa N. ElMeshad, Rania M. Abdelsalam, Mohammed M. Nooh, Muhammad Al-Shorbagy, Gotz Laible
SCIENTIFIC REPORTS
(2017)
Article
Multidisciplinary Sciences
Jingwei Wei, Jisha Antony, Fanli Meng, Paul MacLean, Rebekah Rhind, Gotz Laible, Bjorn Oback
SCIENTIFIC REPORTS
(2017)
Article
Biochemistry & Molecular Biology
Stefan Wagner, Judi McCracken, Sabine Bruszies, Ric Broadhurst, David N. Wells, Bjorn Oback, Juergen Bode, Gotz Laible
MOLECULAR BIOLOGY REPORTS
(2019)
Article
Biotechnology & Applied Microbiology
Swati Jivanji, Chad Harland, Sally Cole, Brigid Brophy, Dorian Garrick, Russell Snell, Mathew Littlejohn, Gotz Laible
Summary: The study did not find any CRISPR-Cas9 associated off-target mutations in the gene-edited cells or calves derived from the gene-edited cell line. Comparison of de novo mutations in two gene-edited calves and three non-edited control calves did not reveal a higher mutation load in either group beyond what was expected from spontaneous mutagenesis. Cell culture and somatic cell nuclear transfer cloning processes were identified as the main sources of contrast in mutational profiles between samples.
Article
Genetics & Heredity
Jingwei Wei, Brigid Brophy, Sally-Ann Cole, Jannis Moormann, Jens Boch, Gotz Laible
Summary: Genome editing allows for targeted introduction of natural sequence variants in cattle breeding, accelerating genetic gain by producing non-mosaic edited embryos with precision genotypes.
FRONTIERS IN GENETICS
(2022)
Article
Nanoscience & Nanotechnology
Muhammed A. Saad, Noha M. Eissa, Mohammed A. Ahmed, Aliaa N. ElMeshad, Goetz Laible, Ahmed S. Attia, Medhat A. Al-Ghobashy, Rania M. Abdelsalam, Muhammad Y. Al-Shorbagy
Summary: A nanoformulation containing rhMBP with RTX on its surface (Nano-rhMBP-RTX) was prepared and found to have potential neuro-protective effects on C57BL/6 mice after inducing experimental autoimmune encephalomyelitis (EAE), improving symptoms and histopathological findings.
INTERNATIONAL JOURNAL OF NANOMEDICINE
(2022)
Article
Biochemistry & Molecular Biology
Goetz Laible, Sally Cole, Brigid Brophy, Paul Maclean, Li How Chen, Dan P. Pollock, Lisa Cavacini, Nathalie Fournier, Christophe De Romeuf, Nicholas C. Masiello, William G. Gavin, David N. Wells, Harry M. Meade
Review
Agronomy
Zachariah Mclean, Bjorn Oback, Gotz Laible
FRONTIERS OF AGRICULTURAL SCIENCE AND ENGINEERING
(2020)
Article
Cell & Tissue Engineering
Stefan Wagner, Alison Cullum, David N. Wells, Gotz Laible
CELLULAR REPROGRAMMING
(2017)