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Posts by Rosy Favicchio

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Preparation of placenta-tropic mRNA lipid nanoparticles for pregnancy disorders Nature Protocols - We present a Protocol for the preparation of placenta-tropic mRNA lipid nanoparticles (LNPs), evaluation of the effects of protein adsorption on mRNA transfection in vitro and...

This mRNA LNP formulation presented is effective for treating pre-eclampsia in mouse models of the condition and could be expanded for the treatment of other pregnancy disorders.

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The procedure covers the synthesis and purification of the ionizable lipid excipient, the preparation of mRNA LNPs, LNP transfection to placental trophoblast cells and the biodistribution and mRNA expression in the placentas of time-dated pregnant mice.

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Preparation of placenta-tropic mRNA lipid nanoparticles for pregnancy disorders - Nature Protocols We present a Protocol for the preparation of placenta-tropic mRNA lipid nanoparticles (LNPs), evaluation of the effects of protein adsorption on mRNA transfection in vitro and assessment of LNP biodis...

A protocol for the preparation and evaluation of placenta-tropic mRNA LNPs for treating pregnancy disorders.
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@natprot.nature.com

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Preparation of targeted lipid nanoparticles for precision nucleic acid delivery Nature Protocols - This protocol outlines the steps to prepare functionalized lipid nanoparticles covalently conjugated with whole antibodies or antibody fragments for targeted nucleic acid...

After antibody labeling and LNP conjugation, tLNPs are purified by size exclusion chromatography and characterized ex vivo and in vivo.

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Strain-promoted azide–alkyne cycloaddition (SPAAC) chemistry is used to covalently conjugate dibenzocyclooctyne (DBCO)-labeled antibodies or antibody fragments to LNPs containing azide.

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Preparation of targeted lipid nanoparticles for precision nucleic acid delivery - Nature Protocols This protocol outlines the steps to prepare functionalized lipid nanoparticles covalently conjugated with whole antibodies or antibody fragments for targeted nucleic acid delivery to different tissues...

This protocol offers a versatile approach to the precise targeting of nucleic acid-containing lipid nanoparticles to cell types or tissues, such as liver- and lung-targeting LNPs in vivo and ex vivo human primary T cell-targeting LNPs.
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Oscillatory brain state-dependent stimulation with transcranial magnetic stimulation combined with electroencephalography Nature Protocols - An approach that couples the application of transcranial magnetic stimulation with brain oscillations to improve the effectiveness of long-term plasticity induction.

This protocol covers brain MRI and image segmentation for anatomical modeling and EEG signal reconstruction, validation of the oscillatory signals, oscillatory phase targeting accuracy and the main experiment of phase-coupled real-time EEG– TMS.

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Oscillatory brain state-dependent stimulation with transcranial magnetic stimulation combined with electroencephalography - Nature Protocols An approach that couples the application of transcranial magnetic stimulation with brain oscillations to improve the effectiveness of long-term plasticity induction.

Real-time EEG–TMS enables the timing of TMS pulses relative to target oscillatory brain states with millisecond-precision
@paolobelardinelli.bsky.social
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Single-cell ultra-high-throughput multiplexed chromatin accessibility and gene expression sequencing (SUM-seq) Nature Protocols - A protocol for simultaneous profiling of chromatin accessibility and gene expression in single nuclei that permits sample multiplexing via sample-specific in situ barcoding and...

This workflow permits simultaneous profiling of chromatin accessibility and gene expression in single nuclei from tens to hundreds of samples and up to 1 million nuclei in a single run and is suitable for a wide variety of sample types.

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Single-cell ultra-high-throughput multiplexed chromatin accessibility and gene expression sequencing (SUM-seq) - Nature Protocols A protocol for simultaneous profiling of chromatin accessibility and gene expression in single nuclei that permits sample multiplexing via sample-specific in situ barcoding and high-throughput droplet...

Single-cell ultra-high-throughput multiplexed sequencing leverages a combinatorial indexing strategy in which chromatin and mRNA are indexed in situ using sample-specific barcodes, and encapsulated into microfluidic droplets. @tommytang.bsky.social
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3D hierarchically aligned nanofiber scaffolds promote cell migration for tissue regeneration Nature Protocols - We present a protocol for the alignment of nanofibers within bioscaffolds to facilitate the recruitment and migration of cells to large wounds, thus improving functional and...

the hierarchical alignment of the nanofibers create pathways for the migrating cells, facilitating their rapid and large-scale transportation from the surrounding tissues to the wound.

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3D hierarchically aligned nanofiber scaffolds promote cell migration for tissue regeneration - Nature Protocols We present a protocol for the alignment of nanofibers within bioscaffolds to facilitate the recruitment and migration of cells to large wounds, thus improving functional and morphological accuracy in ...

A protocol for 3D aligned nanofiber scaffolds provides spatial support for the repair of trauma-induced tissue defects
🧪 #materials
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Volumetric DNA microscopy for mapping spatial transcriptomes in three dimensions Nature Protocols - This protocol for encoding spatial information via DNA sequences enables template-free reconstruction of three-dimensional gene expression patterns in intact specimens, thereby...

The method uses dual-scale spatial encoding through anchored rolling circle amplification for short-range proximities and hydrogel-constrained in vitro transcription for long-range associations, enabling volumetric reconstruction that overcomes previous two-dimensional limitations.

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Volumetric DNA microscopy for mapping spatial transcriptomes in three dimensions - Nature Protocols This protocol for encoding spatial information via DNA sequences enables template-free reconstruction of three-dimensional gene expression patterns in intact specimens, thereby overcoming the limitati...

Volumetric DNA microscopy encodes spatial information into DNA sequences rather than photons, enabling template-free reconstruction of 3D morphology and gene expression patterns in intact specimens through computational analysis of sequencing data. @jaweinst.bsky.social
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High-throughput reaction screening using acoustic ejection mass spectrometry Nature Protocols - A bottleneck in high-throughput chemical synthesis is analyzing reaction products. This protocol describes a direct mass spectrometry approach where the loss of a characteristic...

Acoustic ejection mass spectrometry cuts analysis times from minutes to a single second per sample by fragmenting the original sample and thereby avoiding complex workflows which would otherwise need to be customized for each one, separately.

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High-throughput reaction screening using acoustic ejection mass spectrometry - Nature Protocols A bottleneck in high-throughput chemical synthesis is analyzing reaction products. This protocol describes a direct mass spectrometry approach where the loss of a characteristic neutral fragment from ...

A protocol for determining chemical reactions using acoustic droplet ejection mass spectrometry directly from starting materials enables the fast analysis of hundreds of samples.
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Aqueous sonochemical synthesis of covalent organic frameworks Nature Protocols - Covalent organic frameworks (COFs) could be used for diverse applications. The dominant solvothermal method for COF synthesis requires harsh conditions for long periods of time....

The resulting COFs have large surface areas and high crystallinity, making them suitable for many applications, including photocatalysis, chemical sensing and gas sorption. The method has already been used to prepare 62 COFs with diverse linkages and topologies.

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Aqueous sonochemical synthesis of covalent organic frameworks - Nature Protocols Covalent organic frameworks (COFs) could be used for diverse applications. The dominant solvothermal method for COF synthesis requires harsh conditions for long periods of time. This Protocol describe...

Here, a protocol offering a probe-based sonochemical method for rapidly (under 1 h) synthesizing covalent organic frameworks (COFs) in an aqueous environment under ambient atmosphere, avoiding harmful solvents and high-pressure systems.
🧪 #materials
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Preparing and characterizing environmentally aged microplastics Nature Protocols - Microplastics (MPs) are persistent and widely distributed pollutants. Environmental factors cause physical and chemical aging that change their properties. This protocol is a...

The procedure includes a workflow to generate a composite aging index (CAI) for microplastics that integrates weighted metrics from multiple analytical techniques.

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Preparing and characterizing environmentally aged microplastics - Nature Protocols Microplastics (MPs) are persistent and widely distributed pollutants. Environmental factors cause physical and chemical aging that change their properties. This protocol is a systematic workflow for c...

Microplastic aging is a dynamic process that depends on the chemical composition, size and shape of the material, and its environmental conditions. Here, a protocol to characterize aged microplastics in soil, water, air and organic material.
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High-throughput identification of endogenous biomolecular condensates and phase-separating proteins Nature Protocols - This protocol identifies endogenous biomolecular condensates by combining sucrose density gradient centrifugation with quantitative mass spectrometry, enabling proteome-wide,...

The approach is designed to preserve the condensates in their native cellular state and captures stimulus-responsive condensate dynamic shifts. It provides proteome-wide coverage, enabling unbiased discovery of constitutive and inducible condensates.

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Here, a protocol offering a high-throughput strategy to identify endogenous biomolecular condensates and phase-separating proteins by integrating sucrose density gradient centrifugation with quantitative mass spectrometry.
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“I treated that access as something to be quiet about rather than discussed freely with others”

Peter Attia, explaining how to move in philanthropic circles and raise funds.

Desk reject.

🧪 #academicsky

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Rooting for it! 🍀

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Designing and testing CRISPRi-based synthetic gene circuits in plants Nature Protocols - This protocol details the design principles and procedures for building programmable CRISPRi-based gene circuits, and for testing individual components and the assembled circuits...

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CRISPRi-based circuits can integrate different signals to achieve programmable gene expression in plants, while the high-throughput protoplast-based assay permits rapid and scalable testing of circuit components across different species.

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The procedure covers engineered CRISPRi genetic circuits in plants, and outlines the isolation of protoplasts from different plant species and tissues, and for testing genetic parts and gene circuits using a plate-based, high-throughput, dual luciferase protoplast transfection assay.

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Designing and testing CRISPRi-based synthetic gene circuits in plants - Nature Protocols This protocol details the design principles and procedures for building programmable CRISPRi-based gene circuits, and for testing individual components and the assembled circuits in various plant spec...

Where is the #Arabidopsis community on @bsky.app? Any #Brassica fans? See our recent protocol covering design principles and considerations for building and testing programmable and reversible CRISPRi-based gene circuits in plants.
🧪 #plantscience @natprot.nature.com @ryanlister.bsky.social

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Transsynaptic tracing techniques to interrogate neuronal connectivity of glioblastomas Nature Protocols - This protocol details transsynaptic tracing approaches using retrograde and anterograde viral tracers to map and manipulate neuron–tumor circuits in xenografts, brain...

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The procedure covers diverse experimental models to study neuron–GBM interactions, including the generation of GBO-cortical organoid assembloids, the transplantation of GBM cells into mouse brains and the co-cultures of GBOs with human primary brain tissue surgical slices.

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