Article
Biotechnology & Applied Microbiology
Gang Zhang, Shigang Yao, Yunan Liu, Hailing Fang, Yuqin Song, Chao Wang, Dawei Wei, Jie Feng
Summary: In this study, we identified 530 novel tyrosine-type integrases from bacterial genomic islands. These integrases belong to a distinct evolutionary clade and are affiliated with new integrase groups. The integration functions of six representative novel integrases were validated in a bacterial model system, and their expression patterns were confirmed in Escherichia coli. This study revealed site-specific recombination functions of novel integrases and expands the toolbox for genetic engineering.
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Olli Huhtinen, Rune Salbo, Urpo Lamminmaki, Stuart Prince
Summary: Mammalian display allows for the selection of favorable antibodies from a large antibody library displayed on mammalian cells. A new mammalian display platform was constructed and validated, using the Flp-In CHO cell line. This platform demonstrated the ability to select favorable antibodies and improve the biophysical properties of bococizumab.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2023)
Article
Multidisciplinary Sciences
Jun-Yi Wang, Yue-Yang Cao, Ya-Nan Chen, Xiao-Le Wu, Bo-Tao He, Si-Yu Zhu, Xiao Zhou, Yi Wu, Bing-Zhi Li, Ying-Jin Yuan
Summary: Site-specific recombination systems (SRSs) play an important role in synthetic biology research. Nondirectional SRSs can randomly trigger genome recombination, and in this study, 6 new nondirectional SRSs were designed and successfully generated random recombination in Saccharomyces cerevisiae. These SRSs can independently initiate large-scale gene recombination in different regions of the genome, and they exhibit stability in different cell growth stages and chromosome structures.
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
Agriculture, Dairy & Animal Science
Soo-Young Yum, Woojae Choi, Seokjoong Kim, Goo Jang, Okjae Koo
Summary: Gene integration at site-specific loci, such as pAAVS1, was achieved in porcine fibroblasts using TALEN and CRISPR/Cas9. The efficiency of CRISPR/Cas9 was found to be higher than TALEN. Recombinase-mediated cassette exchange (RMCE) was successfully induced to exchange transgenes at the pAAVS1 locus, demonstrating the usefulness of this technology for porcine transgenesis studies.
ANIMAL BIOTECHNOLOGY
(2023)
Article
Genetics & Heredity
Fang-Jung Yang, Chiao-Nung Chen, Tiffany Chang, Ting-Wei Cheng, Ni-Chen Chang, Chia-Yi Kao, Chih-Chi Lee, Yu-Ching Huang, Jung-Chen Hsu, Jengyi Li, Meiyeh J. Lu, Shih-Peng Chan, John Wang
Summary: In this study, the phiC31 integrase system was adapted for use in C. elegans, allowing for precise single-copy insertion of large DNA constructs and generation of inversions. Integration of genes up to 33.4 kb in size was achieved by injecting a plasmid, and a dual-color fluorescent reporter system for studying post-transcriptional regulation of mRNA was established.
Article
Microbiology
Helena Leinweber, Raphael N. Sieber, Jesper Larsen, Marc Stegger, Hanne Ingmer
Summary: The study revealed that phages carrying virulence factors can adapt and integrate into new strains of Staphylococcus aureus, even those lacking preferred integration sites. This adaptation is facilitated by the promiscuity of the phage-encoded tyrosine recombinase, allowing the phages to increase virulence and zoonotic potential in different host organisms.
Article
Biotechnology & Applied Microbiology
Ke Liu, Gui-Hong Lin, Kun Liu, Yong-Jun Liu, Xin-Yi Tao, Bei Gao, Ming Zhao, Dong-Zhi Wei, Feng-Qing Wang
Summary: In this study, a multiplexed integrase-assisted site-specific recombination (miSSR) method was developed to integrate genes/pathways in Mycolicibacteria for the purpose of constructing cell factories. The method successfully integrated multiple copies of genes using different bacteriophage systems and a reconstructed Xer recombinase system. The results showed that the miSSR system is an effective and convenient tool for engineering the genomes of Mycolicibacteria and other related strains.
SYNTHETIC AND SYSTEMS BIOTECHNOLOGY
(2022)
Article
Chemistry, Multidisciplinary
Ana E. Platero-Prats, Andreas Mavrandonakis, Jian Liu, Zhihengyu Chen, Zhijie Chen, Zhanyong Li, Andrey A. Yakovenko, Leighanne C. Gallington, Joseph T. Hupp, Omar K. Farha, Christopher J. Cramer, Karena W. Chapman
Summary: The study used in situ synchrotron X-ray scattering analysis to evaluate the binding sites for reagent and product molecules near catalytic Ni-oxo clusters in NU-1000 with different surface functionalization, showing a correlation with ethylene hydrogenation activity. The most active catalyst had ethylene reagent molecules binding close to the catalytic clusters only at temperatures near the experimentally observed onset of catalysis, while ethane product molecules favored a different binding location.
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
(2021)
Article
Microbiology
Lauren A. Riley, Irenee C. Payne, Melissa Tumen-Velasquez, Adam M. Guss
Summary: This study developed a system for integrating up to 12 genetic constructs into the E. coli chromosome. Site-specific recombinases were used to catalyze recombination between specific DNA sequences, enabling the integration of heterologous DNA. The utility of this system was demonstrated by the insertion of eight DNA methyltransferases into the E. coli chromosome.
JOURNAL OF BACTERIOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Nina B. Reese, Sherman Ku
Summary: Novel targeted integration systems were developed to achieve high-level expression of recombinant proteins in CHO cells by inserting genes at specific genomic locations, leading to improved expression and productivity. The use of post-integration enrichment significantly increased recombinant protein titer and specific productivity, while maintaining specificity and reproducibility in establishing various cell lines expressing different recombinant protein therapeutics.
CURRENT RESEARCH IN BIOTECHNOLOGY
(2021)
Article
Genetics & Heredity
Leslie A. Mitchell, Laura H. McCulloch, Sudarshan Pinglay, Henri Berger, Nazario Bosco, Ran Brosh, Milica Bulajic, Emily Huang, Megan S. Hogan, James A. Martin, Esteban O. Mazzoni, Teresa Davoli, Matthew T. Maurano, Jef D. Boeke
Summary: Design and large-scale synthesis of DNA have been applied to the functional study of viral and microbial genomes, with new technology development needed for the study of larger mammalian genomes. Challenges include assembling and delivering long DNA sequences. A workflow has been developed for de novo DNA assembly and delivery enabling functional evaluation of mammalian genes at 100kb scale.
Article
Microbiology
Gang Zhang, Qinna Cui, Jianjuan Li, Ruiliang Guo, Sebastien Olivier Leclercq, Lifeng Du, Na Tang, Yuqin Song, Chao Wang, Fangqing Zhao, Jie Feng
Summary: In this study, the mobility of GIsul2 and CR2-sul2 units mediated by integrase was experimentally characterized. The results demonstrate that the integrase of GIsul2 can perform the excision and integration of GIsul2 and CR2-sul2 unit by site-specific recombination between att core sites. The findings suggest a potential and unappreciated role of the GIsul2 integrase family in the dissemination of CR2-sul2 units carrying various resistance determinants.
FRONTIERS IN MICROBIOLOGY
(2022)
Article
Microbiology
Jing Yang, Lina Liu, Yu Feng, Da He, Chengcheng Wang, Zhiyong Zong
Summary: The newly identified mcr-10 plasmid-borne colistin resistance gene was found in a Cronobacter sakazakii isolate. It may be mobilized through site-specific recombination mediated by an integrase of an integrative mobile element.
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY
(2021)
Article
Biochemical Research Methods
Seunghyeon Shin, Su Hyun Kim, Jae Seong Lee, Gyun Min Lee
Summary: The platform developed for targeted integration of transgenes using RMCE and CRISPR/Cas9 shows increased efficiency in dual-landing pad HEK293 cell lines. Accelerating nuclear transport of RMCE components enhanced the efficiency of integrating transgenes at multiple sites without the need for selection or enrichment processes. This streamlined RMCE platform proves to be a versatile tool for facilitating multigene expression at genomic safe harbors in mammalian cell line development.
ACS SYNTHETIC BIOLOGY
(2021)
Article
Genetics & Heredity
Moran Brenner, Lior Lobel, Ilya Borovok, Nadejda Sigal, Anat A. Herskovits
Article
Biology
Inna Rozman Grinberg, Daniel Lundin, Mahmudul Hasan, Mikael Crona, Venkateswara Rao Jonna, Chrishtoph Loderer, Margareta Sahlin, Natalia Markova, Ilya Borovok, Gustav Berggren, Anders Hofer, Derek T. Logan, Britt-Marie Sjoberg
Review
Genetics & Heredity
Gabor Zsurka, Viktoriya Peeva, Alexander Kotlyar, Wolfram S. Kunz
Article
Gastroenterology & Hepatology
Sarit Hoffman, Nathaniel Aviv Cohen, Ian M. Carroll, Hagit Tulchinsky, Ilya Borovok, Iris Dotan, Nitsan Maharshak
JOURNAL OF CROHNS & COLITIS
(2019)
Article
Biochemical Research Methods
Liat Katrivas, Benjamin Kempinski, Kfir Kuchuk, Uri Sivan, Alexander Kotlyar
BIOCONJUGATE CHEMISTRY
(2019)
Article
Biochemistry & Molecular Biology
Inna Rozman Grinberg, Oren Yaniv, Lizett Ortiz de Ora, Ivan Munoz-Gutierrez, Almog Hershko, Oded Livnah, Edward A. Bayer, Ilya Borovok, Felix Frolow, Raphael Lamed, Milana Voronov-Goldman
PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS
(2019)
Article
Nanoscience & Nanotechnology
Galina Proshkina, Sergey Deyev, Anastasiya Ryabova, Francesco Tayanti, Maria Cristina Menziani, Roy Cohen, Liat Katrivas, Alexander Kotlyar
ACS APPLIED MATERIALS & INTERFACES
(2019)
Article
Multidisciplinary Sciences
Tal Argov, Shai Ran Sapir, Anna Pasechnek, Gil Azulay, Olga Stadnyuk, Lev Rabinovich, Nadejda Sigal, Ilya Borovok, Anat A. Herskovits
NATURE COMMUNICATIONS
(2019)
Article
Cell Biology
Anna Pasechnek, Lev Rabinovich, Olga Stadnyuk, Gil Azulay, Jessica Mioduser, Tal Argov, Ilya Borovok, Nadejda Sigal, Anat A. Herskovits
Article
Microbiology
Inna Lysnyansky, Ilya Borovok
Summary: The study identified a prophage-like region in Mycoplasma bovirhinis HAZ141_2 with more than 40% GC content, including a highly transmittable gene cluster that confers high resistance to kanamycin and neomycin.
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY
(2021)
Article
Microbiology
Etai Boichis, Nadejda Sigal, Ilya Borovok, Anat A. Herskovits
Summary: This study identified a conserved locus containing XRE regulators, a metzincin protease, and a lipoprotein structurally similar to a TIMP-family metzincin inhibitor, which play a crucial role in Listeria monocytogenes life cycle. The XRE regulators act as a regulatory switch that influences the expression of metzincin and TIMP-like genes, affecting Lm morphology and virion release. These proteins also modulate the cell wall structure and may hold promise in combating antibiotic resistant bacterial pathogens.
Article
Infectious Diseases
Inna Lysnyansky, Ilya Borovok
Summary: This study characterized recombination events and their consequences within the aadE*-sat4-aphA-3 containing region of the 54 kb GC-rich prophage region in Mycoplasma bovirhinis HAZ141_2. Recombination events including inversions via 155 and 197 bp repeats and the excision of the aadE*-sat4-aphA-3 cluster were confirmed, with effects on gene expression and antibiotic resistance profiles observed. The study highlights the genetic plasticity of the region and its impact on phenotype.
Article
Oncology
Galina Proshkina, Elena Shramova, Anastasiya Ryabova, Liat Katrivas, Clelia Giannini, Daniele Malpicci, Yael Levi-Kalisman, Sergey Deyev, Alexander Kotlyar
Summary: This study demonstrates a methodology to enhance the solubility of peptide nucleic acid (PNA) and delivers it specifically to cancer cells using designed ankyrin repeat protein (DARPin)-functionalized small multilamellar vesicles (SMVs). The positively charged PNA molecules bind electrostatically to the negatively charged phospholipid membranes, encapsulating the PNA in the vesicles. The DARPin-functionalized SMVs can enter HER2-overexpressing cells and cause cell death by interfering with normal cell functioning.
Article
Biochemical Research Methods
Sergey Deyev, Galina Proshkina, Anastasiya Ryabova, Francesco Tavanti, Maria Cristina Menziani, Gennady Eidelshtein, Gavriel Avishai, Alexander Kotlyar
BIOCONJUGATE CHEMISTRY
(2017)
Article
Pharmacology & Pharmacy
Yaron Dekel, Yossy Machluf, Tal Gefen, Gennady Eidelshtein, Alexander Kotlyar, Yaron Bram, Ehud Shahar, Farah Reslane, Elina Aizenshtein, Jacob Pitcovski
