Golgi-IP, a novel tool for multimodal analysis of Golgi molecular content
By onThe authors develop a method for the rapid capture of intact Golgi via Golgi immunoprecipitation. Using high-resolution mass spectrometry, the approach allows the unbiased characterization of the Golgi proteome, metabolome, and lipidome.
The GBA variant E326K is associated with alpha-synuclein aggregation and lipid droplet accumulation in human cell lines.
By onThe GBA variant E326K is associated with alpha-synuclein aggregation and lipid droplet accumulation in human cell lines (associated with publication 10.1101/2022.06.01.494130).
H9 ES AAVS1-NGN2 FAM134C/A/B-/-;TEX264-/-; PiggyBac-Keima-REEP5
By onES cells modified to produce iNeurons w/o ER-phagy receptor FAM134C/A/B, TEX264, and with Keima-REEP5 reporter. CRISPR/Cas9 used to introduce NEUROG2 construct in AAVS1 locus. Knockouts RETREG1/2/3, TEX264. Transfected with NEUROG2, REEP5, mKeima.
Advances in AAV technology for delivering genetically encoded cargo to the nonhuman primate nervous system
By onModern neuroscience approaches including optogenetics, calcium imaging, and other genetic manipulations have facilitated our ability to dissect specific circuits in rodent models to study their role in neurological disease. These approaches regularly use viral vectors to deliver genetic cargo (e.g., opsins) to specific tissues and genetically-engineered rodents to achieve cell-type specificity. However, the translatability of these rodent models, cross-species validation of identified targets, and translational efficacy of potential therapeutics in larger animal models like nonhuman primates remains difficult due to the lack of efficient primate viral vectors. A refined understanding of the nonhuman primate nervous system promises to deliver insights that can guide the development of treatments for neurological and neurodegenerative diseases. Here, we outline recent advances in the development of adeno-associated viral vectors for optimized use in nonhuman primates. These tools promise to help open new avenues for study in translational neuroscience and further our understanding of the primate brain.
Generation of stable ATG3-mCherry wild-type or mutans HeLa cells with HaloTag-LC3B using lentivirus
By onThis protocol details the generation of stable ATG3-mCherry wild-type or mutant HeLa cells with HaloTag-LC3B using lentivirus.
Luminescent Conjugated Oligothiophenes (LCO) Staining
By onThe protocol describes Luminescent Conjugated Oligothiophenes staining on brain sections.
Nestin-cre;Ercc1-/fl mice
By onPan-neuronal cell types may experience higher levels of internal DNA damage and potential senescence due to various factors.
H9 ES AAVS1-NGN2 FAM134A-/-
By onSummary: H9 ES cells were modified using CRISPR/Cas9 to lack the ER-phagy receptor FAM134A. A NEUROG2 construct was introduced in the AAVS1 locus, creating iNeurons. The cells are derived from human embryonic stem cells at the blastocyst stage.
Whole-cell Patch-Clamp Recordings from Striatal Cholinergic Interneurons in ex vivo Mouse Brain Slices
By onProtocol for patch-clamp recording of mCherry-labelled striatal ChIs from mouse brain slices with added step for labelling astrocytes using SR101.
LRRK1 expression and purification
By onProtein purification protocol for full-length LRRK1 as done by Leschziner and Reck-Peterson Labs. Original protocol by Janice M. Reimer and Yu Xuan Lin.
In vitro synthesis of PE2 (nCas9-MMLV RT fusion) polyA mRNA using T7 RNA polymerase
By onWe describe the preparation of synthetic capped and polyadenylated messenger RNA (mRNA) encoding a Cas9 nickase-reverse transcriptase fusion protein (PE2) for use in genome editing with a method called prime editing.
Crystal structure of Wild-Type PPM1H phosphatase
By onCrystal structure of WT PPM1H phosphatase (as reported in 10.15252/embr.202152675) deposited in the Protein Data Bank with code 7L4J.
Create Mask for Axonal Quantification Analysis with FIJI
By onThis protocol describes how to create a quantified mask from an image of fluorescence-tagged axonal projections using FIJI/ImageJ software.
H9 ES AAVS1-NGN2 TEX264-/-
By onES cells modified using CRISPR/Cas9 lack ER-phagy receptor TEX264. They introduced a NEUROG2 construct in AAVS1 locus, derived from a female human embryonic stem cell at the blastocyst stage.
Constitutive nuclear accumulation of endogenous alpha-synuclein in mice causes motor impairment and cortical dysfunction, independent of protein aggregation
By onThe authors created mice in which endogenous a-synuclein is localized to the nucleus. The mice show motor and GI deficits and motor cortex atrophy, suggesting that chronic nuclear a-synuclein can cause toxic phenotypes independent of its aggregation.
RNA and protein extraction from bulk dissections
By onProtocol for RNA and protein extraction from bulk dissections.
An open-source head-fixation and implant-protection system for mice
By onThis study presents a head-fixation and implant-protection system for mice including a headbar, headhat and head fixation station to facilitate experimental procedures in vivo. All 3D-printing files are open source and readily available and editable