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Transcriptional Knockdown in Pneumococci Using CRISPR Interference

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Book cover Streptococcus pneumoniae

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1968))

Abstract

Sequence-specific knockdown of gene expression using CRISPR interference (CRISPRi) has recently been developed for Streptococcus pneumoniae. By coexpression of a catalytically inactive Cas9-protein (dCas9) and a single guide RNA (sgRNA), CRISPRi can be used to knock down transcription of any gene of interest. Gene specificity is mediated by a 20 bp sequence on the sgRNA, and new genes can be targeted by replacing this 20 bp sequence. Here, a protocol is provided for design of sgRNAs and construction of CRIPSRi strains in S. pneumoniae, based on the vectors published by Liu et al. (Mol Syst Biol 13:931, 2017).

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Author information

Authors and Affiliations

  1. Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Ås, Norway

    Morten Kjos

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  1. Morten Kjos
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Correspondence to Morten Kjos .

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Editors and Affiliations

  1. Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Bioclinicum, Stockholm, Sweden

    Federico Iovino

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Kjos, M. (2019). Transcriptional Knockdown in Pneumococci Using CRISPR Interference. In: Iovino, F. (eds) Streptococcus pneumoniae. Methods in Molecular Biology, vol 1968. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9199-0_8

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  • DOI: https://doi.org/10.1007/978-1-4939-9199-0_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9198-3

  • Online ISBN: 978-1-4939-9199-0

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