
Application of CRISPR interference (CRISPRi) for gene silencing in pathogenic species of Leptospira
Full metadata record
DC Field | Value | Language |
---|---|---|
dc.contributor | Lab. Desenvolvimento de Vacinas | pt_BR |
dc.contributor.author | Fernandes, L. G. V. | pt_BR |
dc.contributor.author | Hornsby, R. L. | pt_BR |
dc.contributor.author | Nascimento, Ana Lúcia Tabet Oller do | pt_BR |
dc.contributor.author | Nally, J. E. | pt_BR |
dc.date.accessioned | 2021-09-17T12:56:36Z | - |
dc.date.available | 2021-09-17T12:56:36Z | - |
dc.date.issued | 2021 | pt_BR |
dc.identifier.citation | Fernandes L.G.V., Hornsby R.L., Nascimento ALTO, Nally J.E.. Application of CRISPR interference (CRISPRi) for gene silencing in pathogenic species of Leptospira. Journal of Visualized Experiments. 2021 Aug;174:e62631. doi:10.3791/62631. | pt_BR |
dc.identifier.uri | https://repositorio.butantan.gov.br/handle/butantan/3933 | - |
dc.description.abstract | Leptospirosis is a global neglected zoonosis, responsible for at least 1 million cases per year and almost 60 thousand deaths. The disease is caused by pathogenic and virulent bacteria of the genus Leptospira, either by direct contact with the bacteria or indirectly by exposure to contaminated water or soil. Domestic and wild animals act as reservoir hosts of infection, shedding leptospires from colonized renal tubules of the kidney, via urine, into the environment. The generation of mutant strains of Leptospira is critical to evaluate and understand pathogenic mechanisms of infection. CRISPR interference (CRISPRi) has proven to be a straightforward, affordable, and specific tool for gene silencing in pathogenic Leptospira. Therefore, the methodological details of obtaining the plasmid constructs containing both dCas9 and guide RNA, delivery of plasmids to Leptospira by conjugation with the E. coli strain β2163, and transconjugant recovery and evaluation, will be described. In addition, the recently described Hornsby-Alt-Nally (HAN) media allows for the relatively rapid isolation and selection of mutant colonies on agar plates. | pt_BR |
dc.description.sponsorship | (FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulo | pt_BR |
dc.format.extent | e62631 | pt_BR |
dc.language.iso | English | pt_BR |
dc.relation.ispartof | Journal of Visualized Experiments | pt_BR |
dc.rights | Open access | pt_BR |
dc.title | Application of CRISPR interference (CRISPRi) for gene silencing in pathogenic species of Leptospira | pt_BR |
dc.type | Article | pt_BR |
dc.rights.license | CC BY-NC-ND | pt_BR |
dc.identifier.doi | 10.3791/62631 | pt_BR |
dc.identifier.url | https://doi.org/10.3791/62631 | pt_BR |
dc.contributor.external | (USDA) United States Department of Agriculture | pt_BR |
dc.identifier.citationvolume | 174 | pt_BR |
dc.relation.ispartofabbreviated | JoVE | pt_BR |
dc.identifier.citationabnt | v. 174, e62631, ago. 2021 | pt_BR |
dc.identifier.citationvancouver | 2021 Aug;174:e62631 | pt_BR |
dc.contributor.butantan | Nascimento, Ana Lúcia Tabet Oller do|:Pesquisador|:Laboratório de Desenvolvimento de Vacinas | pt_BR |
dc.sponsorship.butantan | Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2014/50981-0 | pt_BR |
dc.sponsorship.butantan | Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2017/06731-8 | pt_BR |
dc.sponsorship.butantan | Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦ 2019/20302-8 | pt_BR |
dc.identifier.bvscc | BR78.1 | pt_BR |
dc.identifier.bvsdb | IBProd | pt_BR |
dc.description.dbindexed | Yes | pt_BR |
item.fulltext | Com Texto completo | - |
item.grantfulltext | open | - |
item.languageiso639-1 | English | - |
item.openairetype | Article | - |
crisitem.journal.journalissn | #PLACEHOLDER_PARENT_METADATA_VALUE# | - |
crisitem.journal.journaleissn | #PLACEHOLDER_PARENT_METADATA_VALUE# | - |
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crisitem.author.dept | #PLACEHOLDER_PARENT_METADATA_VALUE# | - |
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crisitem.author.orcid | #PLACEHOLDER_PARENT_METADATA_VALUE# | - |
crisitem.author.orcid | #PLACEHOLDER_PARENT_METADATA_VALUE# | - |
crisitem.author.orcid | 0000-0003-4851-0870 | - |
crisitem.author.orcid | #PLACEHOLDER_PARENT_METADATA_VALUE# | - |
crisitem.author.parentorg | #PLACEHOLDER_PARENT_METADATA_VALUE# | - |
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crisitem.author.parentorg | #PLACEHOLDER_PARENT_METADATA_VALUE# | - |
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