...Educational Resources Molecular Biology Reference Molecular Biology Reference Jump To: Molecular Biology Basics...Addgene's molecular biology reference guide, including information on molecular genetics, plasmids, sequencing... molecule is known as DNA or molecular cloning . The process of molecular cloning enabled scientists to...Basics Origins of Molecular Genetics Plasmid Elements Working with Plasmids Introduction Types of Plasmids...Webpage and Blog References Protocols Origins of Molecular Genetics The concept of genes as carriers of phenotypic...These findings culminated in the central dogma of molecular biology, that proteins are translated from RNA...chemistry enable isolation and purification of cellular components, such as DNA, but practically this ...

...both Cas12a and Cas3. Similarly, some Acr proteins are specific to a particular species of CRISPR enzymes... Browse Plasmids: Prime Edit Large Scale Edits CRISPR can generate large-scale genetic changes, including...Cas13a-based molecular detection platform. Based on this non-specific cleavage, a Cas13a-based molecular detection...CRISPR Guide Overview CRISPR Basics Small Edits Large Edits Genome-Wide Screening Cas9 Fusion Tools Cas9...techniques are referred to as CRISPR ( C lustered R egularly I nterspaced S hort P alindromic R epeat) technologies...and adaptability, CRISPR rapidly became the most popular genome engineering approach. Figure 1: Overview...knockout or gene edit; or using Cas9 to generate large genomic deletions or inversions with two gRNA sites...

... Molecular Biology Reference Choosing a Molecular Cloning Technique Molecular Cloning Techniques...Addgene's reference guide on molecular cloning techniques, including information on seven commonly used... Ligation Independent Cloning Oligo Stitching Molecular cloning or the creation of recombinant DNA is ...used in scientific research and discovery. With molecular cloning scientists can amplify and manipulate ...following guide will highlight several of the most popular cloning methods used to create recombinant DNA....Cloning Restriction enzyme (endonuclease) based molecular cloning is the "classic" cloning method, and for... for many reasons, remains one of the most popular today. Restriction enzymes, which are naturally produced...

...and CIB1 450 LARIAT Inhibits protein function by reversibly sequestering targets into large, optically ...Goshen I, Thompson KR, Deisseroth K. 2010 Molecular and cellular approaches for diversifying and extending...cells (frequently neurons) and their governing biomolecular processes. The tools and technologies developed...utilize light to detect, measure, and control molecular signals and cells in order to understand their... Sensors are genetically-encoded reporters of molecular signals; e.g., calcium indicators In this overview...optogenetics tool - they typically allow the fast depolarization of neurons upon exposure to light through direct... trafficks well to the membrane. Excitatory (Depolarizing) ChR Variants. The optogenetic toolbox has been...

...replication as it activates the necessary cellular machinery to produce large amounts of rAAV vectors. The E1 gene...a double-stranded circular episome which can concatemerize, producing high molecular weight structures...retinal disease, spinal muscular atrophy, hemophilia A and B, and Duchenne muscular dystrophy. Many clinical...cloning and working with plasmids, visit Addgene’s Molecular Biology Reference . Production The three production...distributing widely throughout the organism to target large or diffused biological structures. Specifically,...visualize connections between neurons within the brain. Large-scale examples of mapping the brain through neuronal...researchers are used to detect, measure, and control molecular signals of cells (commonly neurons) in response...

...usually VSV-G due to wide infectivity) VSV-G (vesicular stomatitis virus G protein) is the most common...First-generation First-generation lentiviral plasmids are largely defunct due to their lack of safety improvements...on different plasmids, the viral genome remains largely intact. First-generation plasmids include (Figure...cloning and working with plasmids, visit Addgene’s Molecular Biology Reference . Production After cloning, ... shaping integration site preferences. A major cellular determinant is LEDGF (also known as PSIP1 or p75...cells Chimeric envelope genes have also gained popularity to further enhance both infectivity and specificity... dictate the target cell population and reduce cellular toxicity. Common Uses of Lentiviral Vectors Due...

...antigen receptor T cell (CAR-T) therapy is a particularly popular application of gamma-retroviral vectors.... usually VSV-G due to wide infectivity VSV-G (vesicular stomatitis virus G protein) is the most common...cloning and working with plasmids, visit Addgene’s Molecular Biology Reference . Production Using HEK293T Gamma-retroviral...embryonic kidney 293T (HEK293T) cells (Figure 3A), similar to lentiviral packaging methods . The three plasmids...packaging cell line. For example, Phoenix™ is a popular second-generation gamma-retroviral packaging cell...often used to make stable cell lines. Like their popular lentiviral counterparts, gamma-retroviral vectors...Addgene’s protocol is for lentiviral vectors, but a similar method can be applied when using gamma-retroviral...

...vectors, rAdV vectors can provide a large transgene packaging capacity. Large protein-coding genes and complex... First-generation rAdV vectors remain the most popular choice due to their well-established methods, relative...cloning and working with plasmids, visit Addgene’s Molecular Biology Reference . AdEasy™ AdEasy™, developed...developed by Bert Vogelstein’s lab , is the most popular method for creating first-generation rAdV vectors, which...within the nucleus, normally as double-stranded circular episomes. Due to this, and to the strong host’...shorter period of time or when needing to package larger cargo. CRISPR-Cas9 Delivery As compared to other... and third-generation vectors. This capacity is large enough to carry the Cas9 gene and a gRNA expression...

...different reagents that are available for your particular experiment. Select Your Desired Genetic Manipulation...available to carry out your experiment in your particular model organism. There may not be a perfect plasmid...Knockout Permanently disrupt gene function in a particular cell type or organism without a specific preferred... a specific user-defined sequence change in a particular gene, such as generating a point mutation or ...or Interfere (CRISPRi) Reduce expression of a particular gene(s) without permanently modifying the genome...alleles will not contain the desired edit, as a large percentage of DSBs are still repaired by NHEJ. How... and gel electrophoresis (for HDR or NHEJ): For large deletions or insertions, the region of interest ...

...Assays Selection Validation Affinity reagents are molecular tools used in the lab to specifically and tightly...above are used to create hybridomas, which produce large amounts of a single antibody with little variation...antibody is made up of four protein chains: two large heavy chains and two small light chains. These four...In IHC, tissue sections are assayed and the extracellular matrix remains intact. The signaling molecule...enzyme. In ICC, cells have most or all of their extracellular matrix removed. Again, the signaling molecule... fluorophore colors in flow cytometry, but the largest panels used currently are in the mid-forties. When...conjugate or looking for antibodies validated for a similar assay type) and search again. After filtering, ...

...educational resources cover a wide range of topics in molecular biology, laboratory research, and career development...eBooks Download any of our eBooks to learn about popular topics related to our collections. Titles include...plasmids in Addgene’s repository , and related molecular biology techniques. Topics include: Genome Engineering...Viral Vectors Plasmid Technology Career Readiness Molecular Biology Learn about plasmids, sequencing, vectors..., and restriction enzymes to assist with your molecular biology research. Topics include: Antibiotic Concentrations...is a Plasmid? Genetic Code Strain Information Molecular Cloning Techniques Protocols Find written and ...protocols for a wide range of applications in molecular biology, plasmid cloning, and virus, including...

... Chemogenetic tools are actuators for specific cellular pathways, receptors, or ion channels targeted ... on G-protein coupled receptors, or GPCRs. The largest class of cell surface receptors, GPCRs are seven-pass...stimulating phospholipase C, which releases intracellular calcium stores. There are currently three Gq...and activate neuronal signaling by increasing intracellular cAMP concentrations. There is currently one ... was created by replacing the corresponding intracellular region of a turkey erythrocyte β-adrenergic ...muscarinic-receptor based DREADDs. Compound 21 has similar potency as CNO, while DCZ is more potent than either...targeted to specific tissues, cell types or even subcellular regions of a neuron. Cell-specific expression...

...Collection of guides that describe how popular scientific techniques work...activity. Read More CRISPR Class 2 C lustered R egularly I nterspaced S hort P alindromic R epeat (CRISPR...cells (frequently neurons) and their governing biomolecular processes. The tools and technologies developed...utilize light to detect, measure, and control molecular signals and cells in order to understand their...

...designed to cover only a subset of genes, or very large (e.g. the Toronto KnockOut (TKO) library from the... of cells as plasmids in the TKO library. Using large numbers of cells minimizes the effects of random... out our blog posts featuring some of our most popular libraries! Genome-wide Screening Using CRISPR/Cas9... CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens New Tool for Lineage Tracing...

...Addgene's molecular biology reference material covering promoters, transcription and the transcription... Plasmid Reference Molecular Biology Reference Promoters Promoters Definition A promoter is...gRNAs. Resources Addgene's blog , including our popular Plasmids 101 series , covers topics ranging from...plasmid technologies and research to overviews of molecular biology basics and plasmid components. Plasmids...

... Plasmid Reference Molecular Biology Reference Sequencing Primers Sequencing...reference information, please consult Addgene's Molecular Biology Reference Page . All listed primers are...

...September 2023 | Fourth Edition About Based on our popular Plasmids 101 blog series, this eBook covers everything...