Vector Design and Cloning
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Streamlined custom cloning with instant pricing and comprehensive downstream services, including plasmid preparation, virus packaging, library construction, and cell line engineering, all with rigorous QC.
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Featured Citations
Large language models generate functional protein sequences across diverse families
Ali Madani, et al.
Nature Biotechnology, 2023, doi: 10.1038/s41587-022-01618-2
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Abstract: Generating artificial protein sequences using artificial intelligence could enable breakthrough solutions for biomedical and environmental challenges. Viewing amino acid sequences as a language, we demonstrate that a deep learning-based language model can generate functional artificial protein sequences across large protein families, akin to generating grammatically and semantically correct natural language sentences on diverse topics. Our protein language model is trained by simply learning to predict the next amino acid for over 280 million protein sequences from thousands of protein families, without explicit biophysical modeling. We experimentally evaluate model-generated artificial proteins fine-tuned to five distinct antibacterial lysozyme families. Artificial proteins show similar activities and catalytic efficiencies as representative natural lysozymes, including hen egg white lysozyme, while maintaining activity with as low as 31.4% identity to any known naturally-evolved protein. The X-ray crystal structure of an enzymatically active artificial protein recapitulates the conserved fold and positioning of active site residues found in natural proteins. We show our language model’s ability to be adapted to different protein families by accurately predicting the functionality of artificial chorismate mutase and malate dehydrogenase proteins. These results indicate that neural language models successfully perform artificial protein generation across protein families and may prove to be a tool to shortcut evolution.
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Surficial nano-deposition locoregionally yielding bactericidal super CAR-macrophages expedites periprosthetic osseointegration
Ziyang Li, et al.
Science Advances, 2023, doi: 10.1126/sciadv.adg3365
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Abstract: Tracking and eradicating Staphylococcus aureus in the periprosthetic microenvironment are critical for preventing periprosthetic joint infection (PJI), yet effective strategies remain elusive. Here, we report an implant nanoparticle coating that locoregionally yields bactericidal super chimeric antigen receptor macrophages (CAR-MΦs) to prevent PJI. We demonstrate that the plasmid-laden nanoparticle from the coating can introduce S. aureus-targeted CAR genes and caspase-11 short hairpin RNA (CASP11 shRNA) into macrophage nuclei to generate super CAR-MΦs in mouse models. CASP11 shRNA allowed mitochondria to be recruited around phagosomes containing phagocytosed bacteria to deliver mitochondria-generated bactericidal reactive oxygen species. These super CAR-MΦs targeted and eradicated S. aureus and conferred robust bactericidal immunologic activity at the bone-implant interface. Furthermore, the coating biodegradability precisely matched the bone regeneration process, achieving satisfactory osteogenesis. Overall, our work establishes a locoregional treatment strategy for priming macrophage-specific bactericidal immunity with broad application in patients suffering from multidrug-resistant bacterial infection.
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Epicardioid single-cell genomics uncovers principles of human epicardium biology in heart development and disease
Anna B Meier, et al.
Nature Biotechnology, 2023, doi: 10.1038/s41587-023-01718-7
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Abstract: The epicardium, the mesothelial envelope of the vertebrate heart, is the source of multiple cardiac cell lineages during embryonic development and provides signals that are essential to myocardial growth and repair. Here we generate self-organizing human pluripotent stem cell-derived epicardioids that display retinoic acid-dependent morphological, molecular and functional patterning of the epicardium and myocardium typical of the left ventricular wall. By combining lineage tracing, single-cell transcriptomics and chromatin accessibility profiling, we describe the specification and differentiation process of different cell lineages in epicardioids and draw comparisons to human fetal development at the transcriptional and morphological levels. We then use epicardioids to investigate the functional cross-talk between cardiac cell types, gaining new insights into the role of IGF2/IGF1R and NRP2 signaling in human cardiogenesis. Finally, we show that epicardioids mimic the multicellular pathogenesis of congenital or stress-induced hypertrophy and fibrotic remodeling. As such, epicardioids offer a unique testing ground of epicardial activity in heart development, disease and regeneration.
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Educational Resources
Shine Bright, Little Cell: A Comprehensive Guide to Choosing the Right Reporter Gene
Keywords: Reporter genes, fluorescent reporters, luciferase assay
Discover primary reporter types and navigate the roadmap to find whether fluorescent reporters, luciferase, or beta-galactosidase is the best option to detect and measure your gene or element of interest.
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Plasmids Crash Course: Many Hosts, One Plasmid Guest
Keywords: Viral vector, origin of replication, promoter
Learn about the components of a plasmid necessary for gene expression, including promoters, ORFs, polyA signals, selection markers, origins of replication, and antibiotic resistance genes.
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Me First! How Do I Optimize a Vector with Multiple Genes?
Keywords: Bicistronic vector, 2A peptide, linker
This guide provides key considerations and troubleshooting tips for constructing vectors with multiple genes, including which linker to use and the importance of optimizing order.
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My Favorite Cloning Castle
Keywords: PCR, restriction enzymes, ligase
Dive deep into the different steps, strategies, and tricks of gene cloning including different vector backbones, plasmids, PCR, insert types, and gene delivery that are highly adaptable for multiple research scenarios.
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