Genetic tools for the development of recombinant lactic acid bacteria

Wu, Jiapeng; Xin, Yongping; Kong, Jing; Guo, Tingting

doi:10.1186/s12934-021-01607-1

Cited by 22 publications

(23 citation statements)

References 85 publications

(116 reference statements)

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“…Any genome editing procedure typically begins with transformation, which can be accomplished either naturally or artificially by inducing natural competence, or by disrupting the cell membrane with chemicals, electricity, or other forms of disruption [ 1 ]. Natural competence has been observed in several LAB strains [ 14 , 76 , 77 ]. Competence for natural DNA transformation is induced in response to signalling peptides called competence pheromones [ 77 ].…”

Section: Genetic Design Tools Used To Engineer Probioticsmentioning

confidence: 99%

“…Natural competence has been observed in several LAB strains [ 14 , 76 , 77 ]. Competence for natural DNA transformation is induced in response to signalling peptides called competence pheromones [ 77 ]. Uptake of foreign DNA by competent cells typically begins with interaction of the cells with double-stranded DNA in their surroundings.…”

Section: Genetic Design Tools Used To Engineer Probioticsmentioning

confidence: 99%

“…This DNA is translocated into the cytoplasm as a single strand. Upon entry, the single strand is bound by proteins such as the recombination protein (RecA) and the DNA processing protein (DprA), and then it is directly integrated into the genome [ 77 ]. The mechanisms of activating natural competence are not well understood in most LAB.…”

Section: Genetic Design Tools Used To Engineer Probioticsmentioning

confidence: 99%

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Recent advances in genetic tools for engineering probiotic lactic acid bacteria

et al. 2023

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Synthetic biology has grown exponentially in the last few years, with a variety of biological applications. One of the emerging applications of synthetic biology is to exploit the link between microorganisms, biologics, and human health. To exploit this link, it is critical to select effective synthetic biology tools for use in appropriate microorganisms that would address unmet needs in human health through the development of new game-changing applications and by complementing existing technological capabilities. Lactic acid bacteria (LAB) are considered appropriate chassis organisms that can be genetically engineered for therapeutic and industrial applications. Here, we have reviewed comprehensively various synthetic biology techniques for engineering probiotic LAB strains, such as clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 mediated genome editing , homologous recombination, and recombineering. In addition, we also discussed heterologous protein expression systems used in engineering probiotic LAB. By combining computational biology with genetic engineering, there is a lot of potential to develop next-generation synthetic LAB with capabilities to address bottlenecks in industrial scale-up and complex biologics production. Recently, we started working on Lactochassis project where we aim to develop next generation synthetic LAB for biomedical application.

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Section: Genetic Design Tools Used To Engineer Probioticsmentioning

confidence: 99%

Section: Genetic Design Tools Used To Engineer Probioticsmentioning

confidence: 99%

Section: Genetic Design Tools Used To Engineer Probioticsmentioning

confidence: 99%

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Recent advances in genetic tools for engineering probiotic lactic acid bacteria

et al. 2023

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“…However, the mechanisms behind the majority of these interactions have not yet been explored, and only a small number of Gram-positive bacteria have successfully been genetically manipulated. A combination of genome sequence information and insertion mutagenesis will enable the roles of specific genes in host interactions to be confirmed, while genetic engineering provides many new opportunities for the development of such bacteria as therapeutic agents [8,9] . The Roseburia genus is anaerobic Gram-positive bacteria belonging to the Lachnospiraceae family of Firmicutes, and is closely related to Eubacterium rectale [10] .…”

Section: Genetic Manipulation Of Gram-positive Gut Bacteriamentioning

confidence: 99%

Establishing genetic manipulation for novel strains of human gut bacteria

Sheridan¹,

Odat²,

Scott³

2023

Microbiome Res Rep

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Recent years have seen the development of high-accuracy and high-throughput genetic manipulation techniques, which have greatly improved our understanding of genetically tractable microbes. However, challenges remain in establishing genetic manipulation techniques in novel organisms, owing largely to exogenous DNA defence mechanisms, lack of selectable markers, lack of efficient methods to introduce exogenous DNA and an inability of genetic vectors to replicate in their new host. In this review, we describe some of the techniques that are available for genetic manipulation of novel microorganisms. While many reviews exist that focus on the final step in genetic manipulation, the editing of recipient DNA, we particularly focus on the first step in this process, the transfer of exogenous DNA into a strain of interest. Examples illustrating the use of these techniques are provided for a selection of human gut bacteria in which genetic tractability has been established, such as Bifidobacterium, Bacteroides and Roseburia. Ultimately, this review aims to provide an information source for researchers interested in developing genetic manipulation techniques for novel bacterial strains, particularly those of the human gut microbiota.

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“…Remarkably, when selection marker or CRISPR editing alleles are not applicable, transposition is an optimal choice for integrating target genes into the bacterial genome ( Jiang et al, 2013 ; Vo et al, 2021 ). To date, the generally used transposases mainly include sleeping beauty, piggyBac ( Tschorn et al, 2020 ), Tol2 ( Ni et al, 2016 ), Tn5 ( Balasubramanian et al, 2021 ; Xu et al, 2021 ), Tn7 ( Kaczmarska et al, 2022 ), and ICEBs1 ( Ni et al, 2016 ; Peters et al, 2019 ; Strecker et al, 2019 ; Wu et al, 2021 ), all of which possess the ability to integrate small or large DNA fragments into bacterial genomes ( Figure 2 ). However, the transposition efficiency slightly decreases as the size of the inserted DNA fragments increases ( Kowalczykowski, 2015 ).…”

Section: Microbial Genetic Engineeringmentioning

confidence: 99%

Genetically engineered bacterium: Principles, practices, and prospects

et al. 2022

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Advances in synthetic biology and the clinical application of bacteriotherapy enable the use of genetically engineered bacteria (GEB) to combat various diseases. GEB act as a small ‘machine factory’ in the intestine or other tissues to continuously produce heterologous proteins or molecular compounds and, thus, diagnose or cure disease or work as an adjuvant reagent for disease treatment by regulating the immune system. Although the achievements of GEBs in the treatment or adjuvant therapy of diseases are promising, the practical implementation of this new therapeutic modality remains a grand challenge, especially at the initial stage. In this review, we introduce the development of GEBs and their advantages in disease management, summarize the latest research advances in microbial genetic techniques, and discuss their administration routes, performance indicators and the limitations of GEBs used as platforms for disease management. We also present several examples of GEB applications in the treatment of cancers and metabolic diseases and further highlight their great potential for clinical application in the near future.

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Genetic tools for the development of recombinant lactic acid bacteria

Cited by 22 publications

References 85 publications

Recent advances in genetic tools for engineering probiotic lactic acid bacteria

Recent advances in genetic tools for engineering probiotic lactic acid bacteria

Establishing genetic manipulation for novel strains of human gut bacteria

Genetically engineered bacterium: Principles, practices, and prospects

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