ArticleRNA (New York, N.Y.)2006
Restriction for gene insertion within the Lactococcus lactis Ll.LtrB group II intron.
Article in RNA (New York, N.Y.), 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed.
- Development of a group II intron-based genetic manipulation tool for Streptomyces.Microbial biotechnology · 2024Article
- Engineered acetogenic bacteria as microbial cell factory for diversified biochemicals.Frontiers in bioengineering and biotechnology · 2024Review
- SURE editing: combining oligo-recombineering and programmable insertion/deletion of selection markers to efficiently edit the Mycoplasma pneumoniae genome.Nucleic acids research · 2022Article
- Targetron-Assisted Delivery of Exogenous DNA Sequences intoACS synthetic biology · 2021Article
- Molecular characterization of both transesterification reactions of the group II intron circularization pathway.Nucleic acids research · 2021Article
- Group II Introns Generate Functional Chimeric Relaxase Enzymes with Modified Specificities through Exon Shuffling at Both the RNA and DNA Level.Molecular biology and evolution · 2021Article
- U5 snRNA Interactions With Exons Ensure Splicing Precision.Frontiers in genetics · 2021Article
- Synthetic Biology on Acetogenic Bacteria for Highly Efficient Conversion of C1 Gases to Biochemicals.International journal of molecular sciences · 2020Review
- Article
- Development of a metabolic pathway transfer and genomic integration system for the syngas-fermenting bacteriumBiotechnology for biofuels · 2019Article
- Insights Into Non-coding RNAs as Novel Antimicrobial Drugs.Frontiers in genetics · 2019Review
- Recent horizontal transfer, functional adaptation and dissemination of a bacterial group II intron.BMC evolutionary biology · 2016Article
- Extending CRISPR-Cas9 Technology from Genome Editing to Transcriptional Engineering in the Genus Clostridium.Applied and environmental microbiology · 2016Article
- Circularization pathway of a bacterial group II intron.Nucleic acids research · 2016Article
- Bacterial cellular engineering by genome editing and gene silencing.International journal of molecular sciences · 2014Review
- Biotechnological applications of mobile group II introns and their reverse transcriptases: gene targeting, RNA-seq, and non-coding RNA analysis.Mobile DNA · 2014Article
- A targetron system for gene targeting in thermophiles and its application in Clostridium thermocellum.PloS one · 2013Article
- Novel system for efficient isolation of Clostridium double-crossover allelic exchange mutants enabling markerless chromosomal gene deletions and DNA integration.Applied and environmental microbiology · 2012Article
- Gene targeting in gram-negative bacteria by use of a mobile group II intron ("Targetron") expressed from a broad-host-range vector.Applied and environmental microbiology · 2007Article
- Dispersion of the RmInt1 group II intron in the Sinorhizobium meliloti genome upon acquisition by conjugative transfer.Nucleic acids research · 2007Article
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Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The Ll.LtrB intron, from the low G+C gram-positive bacterium Lactococcus lactis, was the first bacterial group II intron shown to splice and mobilize in vivo. The detailed retrohoming and retrotransposition pathways of Ll.LtrB were studied in both L. lactis and Escherichia coli. This bacterial retroelement has many features that would make it a good gene delivery vector. Here we report that the mobility efficiency of Ll.LtrB expressing LtrA in trans is only slightly affected by the insertion of fragments <100 nucleotides within the loop region of domain IV. In contrast, Ll.LtrB mobility efficiency is drastically decreased by the insertion of foreign sequences >1 kb. We demonstrate that the inhibitory effect caused by the addition of expression cassettes on Ll.LtrB mobility efficiency is not sequence specific, and not due to the expression, or the toxicity, of the cargo genes. Using genetic screens, we demonstrate that in order to maintain intron mobility, the loop region of domain IV, more specifically domain IVb, is by far the best region to insert foreign sequences within Ll.LtrB. Poisoned primer extension and Northern blot analyses reveal that Ll.LtrB constructs harboring cargo sequences splice less efficiently, and show a significant reduction in lariat accumulation in L. lactis. This suggests that cargo-containing Ll.LtrB variants are less stable. These results reveal the potential, yet limitations, of the Ll.LtrB group II intron to be used as a gene delivery vector, and validate the random insertion approach described in this study to create cargo-containing Ll.LtrB variants that are mobile.
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