Bacterial genome editing with CRISPR-Cas9: Taking clostridium beijerinckii as an example

Zhong Tian Zhang, Pablo Jiménez-Bonilla, Seungoh Seo, Ting Lu, Yong Su Jin, Hans P. Blaschek, Yi Wang

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

CRISPR-Cas9 has been explored as a transformative genome engineering tool for many eukaryotic organisms. However, its utilization in bacteria remains limited and ineffective. This chapter, taking Clostridium beijerinckii as an example, describes the use of Streptococcus pyogenes CRISPR-Cas9 system guided by the single chimeric guide RNA (gRNA) for diverse genome-editing purposes, including chromosomal gene deletion, integration, single nucleotide modification, as well as “clean” mutant selection. The general principle is to use CRISPR-Cas9 as an efficient selection tool for the edited mutant (whose CRISPR-Cas9 target site has been disrupted through a homologous recombination event and thus can survive selection) against? the wild type background cells. This protocol is broadly applicable to other microorganisms for genome-editing purposes.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages297-325
Number of pages29
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1772
ISSN (Print)1064-3745

Fingerprint

Clostridium beijerinckii
Clustered Regularly Interspaced Short Palindromic Repeats
Bacterial Genomes
Guide RNA
Streptococcus pyogenes
Homologous Recombination
Gene Deletion
Nucleotides
Genome
Bacteria
Gene Editing

Keywords

  • CRISPR-Cas9
  • Clostridium
  • Genome engineering
  • Homologous recombination
  • Single nucleotide modification (SNM)
  • Synthetic biology

PubMed: MeSH publication types

  • Journal Article
  • Research Support, U.S. Gov't, Non-P.H.S.

Cite this

Zhang, Z. T., Jiménez-Bonilla, P., Seo, S., Lu, T., Jin, Y. S., Blaschek, H. P., & Wang, Y. (2018). Bacterial genome editing with CRISPR-Cas9: Taking clostridium beijerinckii as an example. In Methods in Molecular Biology (pp. 297-325). (Methods in Molecular Biology; Vol. 1772). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7795-6_17

Bacterial genome editing with CRISPR-Cas9 : Taking clostridium beijerinckii as an example. / Zhang, Zhong Tian; Jiménez-Bonilla, Pablo; Seo, Seungoh; Lu, Ting; Jin, Yong Su; Blaschek, Hans P.; Wang, Yi.

Methods in Molecular Biology. Humana Press Inc., 2018. p. 297-325 (Methods in Molecular Biology; Vol. 1772).

Research output: Chapter in Book/Report/Conference proceedingChapter

Zhang, ZT, Jiménez-Bonilla, P, Seo, S, Lu, T, Jin, YS, Blaschek, HP & Wang, Y 2018, Bacterial genome editing with CRISPR-Cas9: Taking clostridium beijerinckii as an example. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1772, Humana Press Inc., pp. 297-325. https://doi.org/10.1007/978-1-4939-7795-6_17
Zhang ZT, Jiménez-Bonilla P, Seo S, Lu T, Jin YS, Blaschek HP et al. Bacterial genome editing with CRISPR-Cas9: Taking clostridium beijerinckii as an example. In Methods in Molecular Biology. Humana Press Inc. 2018. p. 297-325. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-7795-6_17
Zhang, Zhong Tian ; Jiménez-Bonilla, Pablo ; Seo, Seungoh ; Lu, Ting ; Jin, Yong Su ; Blaschek, Hans P. ; Wang, Yi. / Bacterial genome editing with CRISPR-Cas9 : Taking clostridium beijerinckii as an example. Methods in Molecular Biology. Humana Press Inc., 2018. pp. 297-325 (Methods in Molecular Biology).
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