Lactobacillus-mediated RNA interference in nematode

Kuwahara, Ai; Arita, Masashi; Kushiro, Akira; Sakube, Yasuji; Sisido, Masahiko; Ohtsuki, Takashi

doi:10.1016/j.jbiosc.2009.08.002

Cited by 5 publications

(3 citation statements)

References 19 publications

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“…Therefore, further development of the siRNA is required to enhance efficiency for future studies. The 5′‐FAM labeled siRNA uptake data revealed that a low level of siRNA entered the bacteria, therefore, other vectors may be investigated in future for improved transformation efficacy .…”

Section: Discussionmentioning

confidence: 99%

Effect of small interfering RNA against Paracin 1.7 bacteriocin produced byLactobacillus paracaseiHD1-7

Wang

Gao

et al. 2015

J. Basic Microbiol.

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Lactobacillus paracasei HD1-7 (CCTCCM 205015), isolated from Chinese sauerkraut fermentation broth, contains the bacteriocin Paracin 1.7 which possesses broad-spectrum antibacterial activity. The gene-silencing effect of small interfering RNA (siRNA) is a potential strategy for further understanding the mechanism of production of Paracin 1.7 by L. paracasei HD1-7. In this study, the effect of siRNA on the expression of the most important proteins in the production of Paracin 1.7, sensor kinase (prcK) and response regulator (prcR), was investigated. SiRNA were designed against prcK and prcR, and qRT-PCR was performed to examine the expression of prcK and prcR mRNA. The efficacy of siRNA was determined by comparing the level of antimicrobial activity of the strains. qRT-PCR showed that siRNA-K4 and siRNA-K5 significantly inhibited the expression of prcK mRNA, and siRNA-R4 and siRNA-R6 significantly inhibited the expression of prcR mRNA. The proteins levels and antibacterial activities of mutant strains were lower than the original and control groups, respectively. The results demonstrate that siRNA inhibited both mRNA expression and the production of Paracin 1.7 in L. paracasei HD1-7. Targeting of prcK and prcR with siRNA appears to be a novel strategy for researching the mechanism of Paracin 1.7 production by L. paracasei HD1-7.

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Section: Discussionmentioning

confidence: 99%

Effect of small interfering RNA against Paracin 1.7 bacteriocin produced byLactobacillus paracaseiHD1-7

Wang

Gao

et al. 2015

J. Basic Microbiol.

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“…1). The emr promoter was amplified from pUCYIT-T7 [17], a derivative of pUCYIT365N, and joined with FLAG-mRFP1 by linking PCR using three primers: 353-emr-AXN, 355-emr-mRFP and 352-mRFP-NBC (Table 2). The P59 promoter sequence was amplified using three overlapping primers: 275-P59-a, 277-P59-b and 276-P59-c (Table 2).…”

Section: Methodsmentioning

confidence: 99%

Distribution Dynamics of Recombinant Lactobacillus in the Gastrointestinal Tract of Neonatal Rats

et al. 2013

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One approach to deliver therapeutic agents, especially proteins, to the gastro-intestinal (GI) tract is to use commensal bacteria as a carrier. Genus Lactobacillus is an attractive candidate for use in this approach. However, a system for expressing exogenous proteins at a high level has been lacking in Lactobacillus. Moreover, it will be necessary to introduce the recombinant Lactobacillus into the GI tract, ideally by oral administration. Whether orally administered Lactobacillus can reach and reside in the GI tract has not been explored in neonates. In this study, we have examined these issues in neonatal rats. To achieve a high level of protein expression in Lactobacillus, we tested the impact of three promoters and two backbones on protein expression levels using mRFP1, a red fluorescent protein, as a reporter. We found that a combination of an L-lactate dehydrogenase (ldhL) promoter of Lactobacillus sakei with a backbone from pLEM415 yielded the highest level of reporter expression. When this construct was used to transform Lactobacillus casei, Lactobacillus delbrueckii and Lactobacillus acidophilus, high levels of mRFP1 were detected in all these species and colonies of transformed Lactobacillus appeared pink under visible light. To test whether orally administered Lactobacillus can be retained in the GI tract of neonates, we fed the recombinant Lactobacillus casei to neonatal rats. We found that about 3% of the bacteria were retained in the GI tract of the rats at 24 h after oral feeding with more recombinant Lactobacillus in the stomach and small intestine than in the cecum and colon. No mortality was observed throughout this study with Lactobacillus. In contrast, all neonatal rats died within 24 hours after fed with transformed E. coli. Taken together, our results indicate that Lactobacillus has the potential to be used as a vehicle for the delivery of therapeutic agents to neonates.

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“…These intrinsic advantages of the LAB strains present an attractive alternative to the use of other mucosal delivery systems with potential problems caused by vehicles such as liposomes and micro-particles [29][30][31]. LAB strains, therefore, have gradually been recognized as one of the promising delivery vehicles for a range of compounds, such as antigens [32], peptides [33], enzymes [34], immune modulators [35], and gene therapy agents [36]. Notably, the first phase I clinical trial conducted in 2006 demonstrated that the application of genetically modified Lactococcus lactis was an effective method for the interleukin-10 topical delivery that circumvented systemic side effects and allowed long-term treatment of chronic diseases [35].…”

Section: Introductionmentioning

confidence: 99%

The Oral Delivery System of Modified GLP-1 by Probiotics for T2DM

Wang

Guo²,

Mao

et al. 2023

Pharmaceutics

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The glucagon-like peptide-1 (GLP-1) is a peptide with incretin activity and plays an important role in glycemic control as well as the improvement of insulin resistance in type 2 diabetes mellitus (T2DM). However, the short half-life of the native GLP-1 in circulation poses difficulties for clinical practice. To improve the proteolytic stability and delivery properties of GLP-1, a protease-resistant modified GLP-1 (mGLP-1) was constructed with added arginine to ensure the structural integrity of the released mGLP-1 in vivo. The model probiotic Lactobacillus plantarum WCFS1 was chosen as the oral delivery vehicle with controllable endogenous genetic tools driven for mGLP-1 secretory constitutive expression. The feasibility of our design was explored in db/db mice which showed an improvement in diabetic symptoms related to decreased pancreatic glucagon, elevated pancreatic β-cell proportion, and increased insulin sensitivity. In conclusion, this study provides a novel strategy for the oral delivery of mGLP-1 and further probiotic transformation.

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Lactobacillus-mediated RNA interference in nematode

Cited by 5 publications

References 19 publications

Effect of small interfering RNA against Paracin 1.7 bacteriocin produced byLactobacillus paracaseiHD1-7

Effect of small interfering RNA against Paracin 1.7 bacteriocin produced byLactobacillus paracaseiHD1-7

Distribution Dynamics of Recombinant Lactobacillus in the Gastrointestinal Tract of Neonatal Rats

The Oral Delivery System of Modified GLP-1 by Probiotics for T2DM

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