دورية أكاديمية
Genetic manipulation of an Ixodes scapularis cell line.
| العنوان: | Genetic manipulation of an Ixodes scapularis cell line. |
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| المؤلفون: | Singh N; Department of Microbiology and Immunology, School of Medicine, University of Maryland, Baltimore, Maryland, USA., Rolandelli A; Department of Microbiology and Immunology, School of Medicine, University of Maryland, Baltimore, Maryland, USA., O'Neal AJ; Department of Microbiology and Immunology, School of Medicine, University of Maryland, Baltimore, Maryland, USA., Butler LR; Department of Microbiology and Immunology, School of Medicine, University of Maryland, Baltimore, Maryland, USA., Samaddar S; Department of Microbiology and Immunology, School of Medicine, University of Maryland, Baltimore, Maryland, USA., Laukaitis-Yousey HJ; Department of Microbiology and Immunology, School of Medicine, University of Maryland, Baltimore, Maryland, USA., Butnaru M; Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts, USA.; Howard Hughes Medical Institute, Chevy Chase, Maryland, USA., Mohr SE; Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts, USA., Perrimon N; Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts, USA.; Howard Hughes Medical Institute, Chevy Chase, Maryland, USA., Pedra JHF; Department of Microbiology and Immunology, School of Medicine, University of Maryland, Baltimore, Maryland, USA. |
| المصدر: | MBio [mBio] 2024 Mar 13; Vol. 15 (3), pp. e0247923. Date of Electronic Publication: 2024 Feb 21. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Society for Microbiology Country of Publication: United States NLM ID: 101519231 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2150-7511 (Electronic) NLM ISO Abbreviation: mBio Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, D.C. : American Society for Microbiology |
| مواضيع طبية MeSH: | Ixodes* , Borrelia burgdorferi*/genetics , Anaplasma phagocytophilum*/genetics , Rickettsia*, Animals ; Humans ; Cell Line ; Mammals |
| مستخلص: | Although genetic manipulation is one of the hallmarks of model organisms, its applicability to non-model species has remained difficult due to our limited understanding of their fundamental biology. For instance, manipulation of a cell line originated from the black-legged tick Ixodes scapularis, an arthropod that serves as a vector for several human pathogens, has yet to be established. Here, we demonstrate the successful genetic modification of the commonly used tick ISE6 line through ectopic expression and clustered regularly interspaced palindromic repeats [(CRISPR)/CRISPR-associated protein 9 (Cas9)] genome editing. We performed ectopic expression using nucleofection and attained CRISPR-Cas9 editing via homology-dependent recombination. Targeting the E3 ubiquitin ligase x-linked inhibitor of apoptosis ( xiap ) and its substrate p47 led to an alteration in molecular signaling within the immune deficiency network and increased infection of the rickettsial agent Anaplasma phagocytophilum in I. scapularis ISE6 cells. Collectively, our findings complement techniques for the genetic engineering of I. scapularis ticks , which currently limit efficient and scalable molecular genetic screens in vivo .IMPORTANCEGenetic engineering in arachnids has lagged compared to insects, largely because of substantial differences in their biology. This study unveils the implementation of ectopic expression and CRISPR-Cas9 gene editing in a tick cell line. We introduced fluorescently tagged proteins in ISE6 cells and edited its genome via homology-dependent recombination. We ablated the expression of xiap and p47 , two signaling molecules present in the immune deficiency (IMD) pathway of Ixodes scapularis . Impairment of the tick IMD pathway, an analogous network of the tumor necrosis factor receptor in mammals, led to enhanced infection of the rickettsial agent Anaplasma phagocytophilum . Altogether, our findings provide a critical technical resource to the scientific community to enable a deeper understanding of biological circuits in the black-legged tick I. scapularis . Competing Interests: The authors declare no conflict of interest. |
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| معلومات مُعتمدة: | F31 AI152215 United States AI NIAID NIH HHS; T32 AI162579 United States AI NIAID NIH HHS; P01 AI138949 United States AI NIAID NIH HHS; F31 AI167471 United States AI NIAID NIH HHS; R21 AI168592 United States AI NIAID NIH HHS; R21 AI165520 United States AI NIAID NIH HHS; R01 AI116523 United States AI NIAID NIH HHS; R01 AI134696 United States AI NIAID NIH HHS |
| فهرسة مساهمة: | Keywords: Borrelia burgdorferi; Lyme disease; rickettsia; tick-borne diseases; ticks |
| تواريخ الأحداث: | Date Created: 20240221 Date Completed: 20240314 Latest Revision: 20240315 |
| رمز التحديث: | 20240315 |
| مُعرف محوري في PubMed: | PMC10936194 |
| DOI: | 10.1128/mbio.02479-23 |
| PMID: | 38380961 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 2150-7511 |
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| DOI: | 10.1128/mbio.02479-23 |