Article
Immunology
Agata N. Rumianek, Ben Davies, Keith M. Channon, David R. Greaves, Gareth S. D. Purvis
Summary: In this study, a novel transgenic mouse (hCD68-CreERT2) was successfully generated to target macrophages in vivo under continuous tamoxifen induction, without affecting the expression of other myeloid cells. Through mating with the tdTomato variant mouse, the efficiency and specificity of this transgenic mouse were validated.
FRONTIERS IN IMMUNOLOGY
(2022)
Article
Endocrinology & Metabolism
Y. Kitase, M. Prideaux
Summary: This article summarizes the main methods for studying the functions of bone cells, including gene knockout and cell tracing. However, there are concerns about the specificity of the promoters used and the off-target effects on cells within and outside of the bone. A thorough understanding of when and where these promoters are activated will improve the quality of study design and the confidence in data interpretation.
Article
Pediatrics
Qilei Han, Ruling Shen, Kai Li, Jian Fei
Summary: The purpose of this study was to induce the expression of Amphinase in neuroblastoma cell lines and establish a foundation for mechanism study. A loxP-cassette vector was constructed and transfected into neuroblastoma cells. Stable transfection was confirmed by PCR and qPCR. The expression of Amphinase was activated by Cre recombinase, resulting in inhibition of cell proliferation. RNA sequencing analysis revealed the impact of Amphinase on the ER function of neuroblastoma cells. In conclusion, the Cre/loxP system successfully induced the expression of Amphinase and provided a powerful tool for mechanism study.
JOURNAL OF PEDIATRIC SURGERY
(2023)
Article
Immunology
Guan Wang, Cunling Zhang, Hiroto Kambara, Cheryl Dambrot, Xuemei Xie, Li Zhao, Rong Xu, Andrea Oneglia, Fei Liu, Hongbo R. Luo
Summary: This study used TLA sequencing to determine the integration site and flanking sequences of the MRP8-Cre-ires/EGFP transgene, revealing its integration into chromosome 5 of the host mouse genome, leading to a deletion of host genomic sequence. The researchers designed a new genotyping protocol, but found that crossing heterozygous mice did not produce homozygous mice, likely due to disrupted Ap1s1 gene expression resulting in prenatal lethality.
FRONTIERS IN IMMUNOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Leonid A. Ilchuk, Nina Stavskaya, Ekaterina A. Varlamova, Alvina Khamidullina, Victor V. Tatarskiy, Vladislav A. Mogila, Ksenia B. Kolbutova, Sergey A. Bogdan, Alexey M. Sheremetov, Alexandr N. Baulin, Irina A. Filatova, Yulia Yu Silaeva, Maxim A. Filatov, Alexandra Bruter
Summary: This article investigates the efficiency of different induction methods on knockout and finds that efficiency is correlated with the concentration of tamoxifen metabolites. Two specific cases of tamoxifen-induced knockout limitations are described. Increasing tamoxifen dosage and activation duration improves knockout rate in the brain but not in the uterus. Induction of knockout during embryonic development is impossible due to the negative effect of tamoxifen on gestation.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Virology
Nannan Liu, Chenjian Gu, Yang Yang, Zixiang Gao, Ning Kang, Jing Liu, Youhua Xie
Summary: This study reports a method for the generation of circular viral genomes through DNA recombination in cells. It allows for simpler manipulation of the BKV genome and results in higher virus yield.
JOURNAL OF MEDICAL VIROLOGY
(2023)
Article
Entomology
Guangsheng Yuan, Jiaxin Shi, Jiahao Shi, Dandan He, Youliang Li, Juan Du, Chaoying Zou, Langlang Ma, Guangtang Pan, Yaou Shen
Summary: In this study, a Cre/loxP-based strategy was proposed to control the expression of transgenic proteins in green and non-green tissues of genetically modified maize. The results showed that by crossing two different transgenic maize plants, the targeted gene expression was achieved specifically in green tissues, while being blocked in root and kernel tissues. Moreover, the transgenic maize exhibited high resistance against common pests in both laboratory and field experiments. These findings are important for the development of novel genetic engineering techniques and addressing public concerns about the biosafety of genetically modified plants.
JOURNAL OF ECONOMIC ENTOMOLOGY
(2023)
Article
Biochemical Research Methods
Peng-Yang Liu, Gang Li, Chu-Bin Lin, Jun-Jie Wu, Shan Jiang, Feng-Hong Huang, Xia Wan
Summary: By establishing a seamless genome editing system, we successfully enhanced the production of astaxanthin in Schizochytrium sp., broadening the application of this strain.
ACS SYNTHETIC BIOLOGY
(2022)
Article
Gastroenterology & Hepatology
Robert L. Kruse, Mercedes Barzi, Xavier Legras, Francis P. Pankowicz, Nika Furey, Lan Liao, Janming Xu, Beatrice Bissig-Choisat, Betty L. Slagle, Karl-Dimiter Bissig
Summary: This study developed a novel transgenic mouse model that allows for excision of HBV from mouse chromosomes to generate a recombinant circular DNA molecule. The model showed regulation of HBV expression and demonstrated the potential for HBV clearance.
Article
Microbiology
Haiyan Ren, Anqi Yin, Pingping Wu, Huanyu Zhou, Jungang Zhou, Yao Yu, Hong Lu
Summary: In this study, the positions and sequences of centromeres in Kluyveromyces marxianus were determined for the first time, and a Cre-loxP system was successfully established in this species. A K. marxianus strain containing 16 loxPsym sites was constructed, and large-scale chromosomal rearrangements were achieved with a high recombination frequency. These findings provide valuable information and tools for studying chromosomal structures and functions in K. marxianus.
Article
Biochemistry & Molecular Biology
Chen Yang, Jia Ge, Xiaokang Fu, Keming Luo, Changzheng Xu
Summary: The study introduced a novel gene deletion system mediated by dual reproductive cell-specific promoters in transgenic plants, enabling the removal of exogenous genes while maintaining desirable traits. Recombination efficiency was validated through hybridization and self-crossing, with over 59% deletion efficiency achieved in the F1 progeny, suggesting the potential of alleviating biosafety concerns while benefiting from GM crop advantages.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Immunology
Thao Vo, Yogesh Saini
Summary: The Cre-LoxP system is commonly used for recombination of floxed genes, but current macrophage-specific Cre recombinase mouse strains have limitations. This study tested the Mafb-Cre strain, which labels macrophages in various organs. The results show that Mafb-Cre only targets around 40% of alveolar macrophages, regardless of age. Mafb+ macrophages exhibit higher expression of surface markers. Bone marrow-derived macrophages are highly amenable to Cre-LoxP recombination, and Th2 stimulation and ozone exposure promote Mafb-Cre-mediated recombination in alveolar macrophages.
FRONTIERS IN IMMUNOLOGY
(2022)
Review
Immunology
Marc Dalod, Stefanie Scheu
Summary: DCs play a crucial role in antigen presentation and orchestrate information transfer in the immune system. Mouse models provide valuable insights into the different functions of DC types and their activation states. This article provides comprehensive information on selecting mutant mouse strains for DC research and discusses the advantages, limitations, and future perspectives of these models.
EUROPEAN JOURNAL OF IMMUNOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Irena Krueger, Friedrich Reusswig, Kim Juergen Krott, Celina Fabienne Lersch, Martina Spelleken, Margitta Elvers
Summary: The study demonstrates that the mT/mG Cre reporter mouse is a suitable model for real-time visualization of platelets, analysis of cell morphology, and identification of non-recombined platelets. However, a small proportion of leukocytes exhibit mG fluorescence in PF4-Cre positive mice. Therefore, careful evaluation of platelet analysis beyond hemostasis and thrombosis should be conducted when increasing recombination of immune cells.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Microbiology
Wataru Kitagawa, Miyako Hata
Summary: This study developed an effective plasmid-curing method and a simple two-step genome-reduction method using a modified Cre/loxP system. A mutant strain that cured all four plasmids and deleted seven chromosomal regions was successfully obtained in this study. This genetic tool will help develop and improve Rhodococcus strains for various industrial and environmental applications.