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Gene Silencing: Interruption or suppression of the expression of a gene at transcriptional or translational levels.

Gene Silencing with Morpholinos

JoVE 5326

Morpholino-mediated gene silencing is a common technique used to study roles of specific genes during development. Morpholinos inhibit gene expression by hybridizing to complementary mRNAs. Due to their unique chemistry, morpholinos are easy to produce and store, which makes them remarkably cost effective compared to other gene silencing methods.


This video reviews proper…

 Developmental Biology

An Overview of Gene Expression

JoVE 5546

Gene expression is the complex process where a cell uses its genetic information to make functional products. This process is regulated at multiple stages, and any misregulation could lead to diseases such as cancer.

This video highlights important historical discoveries relating to gene expression, including the…

 Genetics

MicroRNAs

JoVE 10801

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA ends have been methylated to prevent degradation, it is exported from the nucleus into the cytoplasm. In the cytoplasm, another endonuclease enzyme, called Dicer, cuts the pre-miRNA into a 21-24 nucleotide-long miRNA duplex. Then, Dicer cleaves one strand of the duplex, releasing a single strand of mature miRNA. The mature miRNA is loaded into a protein complex called RNA-induced silencing complex (RISC), which the miRNA then guides to the complementary region of its target mRNA. The extent of complementary base-pairing between miRNA and 3’ untranslated region of target mRNA determines the gene silencing mechanism. Extensive or near-perfect complementarity causes degradation of mRNA, whereas limited base-pairing inhibits translation. While silencing via mRNA degradation is irreversible, translation inhibition is reversible since stable mRNA can

 Core: Gene Expression

Neuronal Transfection Methods

JoVE 5215

Transfection - the process of transferring genetic material into cells - is a powerful tool for the rapid and efficient manipulation of gene expression in cells. Because this method can be used to silence the expression of specific proteins or to drive the expression of foreign or modified proteins, transfection is an extremely useful tool in the study of the cellular and…

 Neuroscience

DNA Methylation Analysis

JoVE 5550

Methylation at CpG dinucleotides is a chemical modification of DNA hypothesized to play important roles in regulating gene expression. In particular, the methylation of clusters of methylation sites, called “CpG islands”, near promoters and other gene regulatory elements may contribute to the stable silencing of genes, for example, during epigenetic processes…

 Genetics

Zebrafish Microinjection Techniques

JoVE 5130

One of the major advantages to working with zebrafish (Danio rerio) is that their genetics can be easily manipulated by microinjection of early stage embryos. Using this technique, solutions containing genetic material or knockdown constructs are delivered into the blastomeres: the embryonic cells sitting atop the yolk of the newly fertilized egg. Delivery into the cytoplasm is…

 Biology II

RNAi in C. elegans

JoVE 5105

RNA interference (RNAi) is a widely used technique in which double stranded RNA is exogenously introduced into an organism, causing knockdown of a target gene. In the nematode, C. elegans, RNAi is particularly easy and effective because it can be delivered simply by feeding the worms bacteria that express double stranded RNA (dsRNA) that is complementary to a gene of interest. First,…

 Biology I

Chromatin Immunoprecipitation

JoVE 5551

Histones are proteins that help organize DNA in eukaryotic nuclei by serving as “scaffolds” around which DNA can be wrapped, forming a complex called “chromatin”. These proteins can be modified through the addition of chemical groups, and these changes affect gene expression. Researchers use a technique called chromatin immunoprecipitation (ChIP) to …

 Genetics

In ovo Electroporation of Chicken Embryos

JoVE 5156

Electroporation is a technique used in biomedical research that allows for the manipulation of gene expression via the delivery of foreign genetic material into cells. More specifically, in ovo electroporation is performed on early developing chicks (Gallus gallus domesticus) contained within their eggshells. In this procedure, DNA or knockdown constructs are first injected…

 Biology II

An Introduction to Transfection

JoVE 5068

Transfection is the process of inserting genetic material, such as DNA and double stranded RNA, into mammalian cells. The insertion of DNA into a cell enables the expression, or production, of proteins using the cells own machinery, whereas insertion of RNA into a cell is used to down-regulate the production of a specific protein by stopping translation. While the site of action for…

 Basic Methods in Cellular and Molecular Biology

An Introduction to Cellular and Molecular Neuroscience

JoVE 5213

Cellular and molecular neuroscience is one of the newest and fastest growing subdisciplines in neuroscience. By investigating the influences of genes, signaling molecules, and cellular morphology, researchers in this field uncover crucial insights into normal brain development and function, as well as the root causes of many pathological conditions.


 Neuroscience

CRISPR-mediated Loss of Function Analysis in Cerebellar Granule Cells Using In Utero Electroporation-based Gene Transfer

1Division of Molecular Neurogenetics, German Cancer Research Center (DKFZ), DKFZ-ZMBH Alliance, 2Division of Molecular Genetics, German Cancer Research Center (DKFZ), and German Cancer Consortium (DKTK), 3Hopp-Children's Cancer Center at the NCT Heidelberg (KiTZ), 4Division of Pediatric Neurooncology, German Cancer Research Center (DKFZ), and German Cancer Consortium (DKTK), 5Department of Pediatric Hematology and Oncology, Heidelberg University Hospital

JoVE 57311

 Developmental Biology

Ultrahigh Resolution Mouse Optical Coherence Tomography to Aid Intraocular Injection in Retinal Gene Therapy Research

1Research Service, VA Western New York Healthcare System, 2Department of Ophthalmology, (Ross Eye Institute), Jacobs School of Medicine and Biomedical Sciences, University at Buffalo- SUNY, 3Pharmacology/Toxicology, Jacobs School of Medicine and Biomedical Sciences, University at Buffalo- SUNY, 4Physiology/Biophysics, Jacobs School of Medicine and Biomedical Sciences, University at Buffalo- SUNY, 5Neuroscience Program, Jacobs School of Medicine and Biomedical Sciences, University at Buffalo- SUNY, 6The RNA Institute, University at Buffalo- SUNY, 7The SUNY Eye Institute

JoVE 55894

 Medicine

Genetic Engineering of Primary Mouse Intestinal Organoids Using Magnetic Nanoparticle Transduction Viral Vectors for Frozen Sectioning

1Department of Medicine, Division of Hematology, Johns Hopkins University School of Medicine, 2Department of Natural Sciences, School of Arts & Sciences, Lebanese American University, 3Department of Cell Biology, Johns Hopkins University School of Medicine, 4Department of Oncology, Johns Hopkins University School of Medicine, 5Department of Pathology, Johns Hopkins University School of Medicine, 6Institute for Cell Engineering, Johns Hopkins University School of Medicine

JoVE 57040

 Developmental Biology

Lentiviral Vector Platform for the Efficient Delivery of Epigenome-editing Tools into Human Induced Pluripotent Stem Cell-derived Disease Models

1Department of Neurology, Duke University Medical Center, 2Center for Genomic and Computational Biology, Duke University Medical Center, 3Viral Vector Core, Duke University Medical Center, 4Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, Duke University Medical Center

JoVE 59241

 Genetics

Genome-wide RNAi Screening to Identify Host Factors That Modulate Oncolytic Virus Therapy

1Children's Hospital of Eastern Ontario (CHEO) Research Institute, 2Department of Biology, Microbiology and Immunology, University of Ottawa, 3Department of Pediatrics, University of Ottawa, 4Department of Microbiology, Immunology and Infectious Diseases, Cumming School of Medicine, University of Calgary

JoVE 56913

 Cancer Research

A Primary Neuron Culture System for the Study of Herpes Simplex Virus Latency and Reactivation

1Department of Microbiology, New York University School of Medicine, 2Molecular Neurobiology Program, Skirball Institute for Biomolecular Medicine, New York University School of Medicine, 3Department of Otolaryngology, New York University School of Medicine, 4Department of Cell Biology, New York University School of Medicine, 5Department of Physiology and Neuroscience, New York University School of Medicine, 6Department of Psychiatry, New York University School of Medicine, 7Center for Neural Science, New York University School of Medicine

JoVE 3823

 Immunology and Infection

Generation, Amplification, and Titration of Recombinant Respiratory Syncytial Viruses

1UMR 1173 Institut national de la santé et de la recherche médicale (INSERM), Université de Versailles St. Quentin, 2UR892 Institut national de la recherche agronomique (INRA), Unité de virologie et immunologie moléculaires, 3Assistance Publique–Hôpitaux de Paris (AP-HP), Hôpital Ambroise Paré, Laboratoire de Microbiologie

JoVE 59218

 Immunology and Infection

Inducible, Cell Type-Specific Expression in Arabidopsis thaliana Through LhGR-Mediated Trans-Activation

1Department of Developmental Physiology, Centre for Organismal Studies (COS) Heidelberg, 2Department of Cell Biology, Centre for Organismal Studies (COS) Heidelberg, 3Department of Stem Cell Biology, Centre for Organismal Studies (COS) Heidelberg

JoVE 59394

 Genetics
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