Protocol for sparse labeling of murine Purkinje cells by intracerebroventricular AAV injection using a microelectrode holder [0.03%]
小鼠脑室内AAV注射稀疏标记浦肯野细胞的协议（利用微电极夹）
Jordan J Lillibridge,Samantha Vancs,Martin M Riccomagno et al.
Jordan J Lillibridge et al.
Most microinjection techniques require expensive machinery and can be difficult to perform consistently. Here, we present a protocol for long-term, stable gene delivery by adeno-associated virus (AAV) vectors through intracerebroventricular...
Protocol for one-step design of a triple sgRNA-based CRISPR/Cas9 construct for neuronal gene knockout in the mouse brain [0.03%]
一步构建用于小鼠脑神经元基因敲除的三sgRNA载体的协议
Gibson Dowd,Yuki Ogawa
Gibson Dowd
CRISPR-Cas9 enables genome editing through the expression of Cas9 and single guide RNAs (sgRNAs). Here, we present a protocol for designing and constructing the vector expressing three sgRNAs targeting a single gene in the mouse brain. We d...
Protocol for analyzing protein interactomes under starvation in fission yeast using in vivo cross-linking [0.03%]
利用体内交联分析裂殖酵母饥饿条件下的蛋白质相互作用组的方案
Livia Pérez-Hidalgo,Celia Gálvez-Merchán,Javier Encinar Del Dedo et al.
Livia Pérez-Hidalgo et al.
Identifying a protein's interactome can be challenging when studying low-abundance proteins, labile or transient interactions, or cells under starvation conditions. Here, we present a protocol to purify proteins and their interacting partne...
Protocol for isolation of the brain vasculature from the mouse cortex for proteomics [0.03%]
小鼠大脑皮层脑血管的蛋白质组学分离协议
Yazi Huang,Veronica Clementel,Mingzi Zhang et al.
Yazi Huang et al.
The neurovascular unit, composed of brain endothelial cells, pericytes, and astrocytes, regulates molecular exchange between blood and the central nervous system. Here, we present a protocol to isolate brain vessels from the mouse cortex us...
Protocol for in vivo transfection of mRNA-encapsulated lipid nanoparticles in mouse testis [0.03%]
小鼠睾丸中包裹mRNA的脂质纳米颗粒体内转染协议
Liying Wang,Huafang Wei,Yiyao Wu et al.
Liying Wang et al.
Male infertility affects 10%-20% of the population in most countries, with approximately half of the cases attributed to male factors. Here, we present a protocol for in vivo transfection of mRNA-encapsulated lipid nanoparticles (mRNA-LNPs)...
Protocol for mapping gene essentiality changes in yeast using inducible gene deletion mutants [0.03%]
在酵母中使用诱导型基因敲除菌株绘制必需基因变化的协议
Sabine van Schie,Jolanda van Leeuwen
Sabine van Schie
Gene essentiality can differ among genetically distinct individuals. In this protocol, we describe a systematic approach for identifying genes that are essential in a laboratory strain of the budding yeast Saccharomyces cerevisiae but vary ...
Protocol for high-throughput droplet-engineered TIL-retaining tumor organoids in triple-negative breast cancer [0.03%]
三阴性乳腺癌的高通量液滴工程肿瘤类器官保留TIL协议
Xiaolong Zhou,Yu Zhu,Wanlong Wang et al.
Xiaolong Zhou et al.
Here, we present a protocol for generating high-throughput, droplet-engineered tumor organoids from triple-negative breast cancer (TNBC) tissues using the OrgFab® micro-sphere printing system. We describe tumor dissociation, bioink prepara...
Protocol for HIV-1 latency reversal using engineered bacteriophage T4 particles [0.03%]
利用工程噬菌体T4颗粒逆转HIV-1潜伏感染的方案
Himanshu Batra,Venigalla B Rao
Himanshu Batra
A major barrier to curing HIV-1 is the latent viral reservoir in resting CD4+ T cells. Here, we present a protocol for preparing CD4-DARPin (designed ankyrin repeat protein)-T4 nanoparticles and their application to HIV-1 latency reversal i...
Isotope-free mapping of protein-RNA interactions at single-nucleotide resolution by iCLIP3 [0.03%]
无同位素的iCLIP3技术：单核苷酸分辨率下的蛋白质-RNA相互作用图谱绘制
Vladimir Despic,Melina Klostermann,Anna Orekhova et al.
Vladimir Despic et al.
Individual-nucleotide resolution UV crosslinking and immunoprecipitation (iCLIP) enables transcriptome-wide mapping of RNA-binding protein (RBP)-RNA interactions. Here, we present iCLIP version 3 (iCLIP3), a streamlined protocol optimized f...
Romuald Binet,Claire F Pearson
Romuald Binet
This overview provides novices with the most important factors to consider when planning to set up a germ-free facility to house sterile mice. This includes deciding what facility model is most appropriate for the required project, which wi...