...Each system also includes Tn7-like transposase components. Type I (generally) includes tnsA , tnsB , tnsC...applications to increasingly complex functions, including the ability to selectively activate/repress target...carried out by Cas9. Multiplexing applications include editing multiple genes at once; using dual nickases...increased fidelity enzymes, and their mechanisms, include: eSpCas9(1.1) - weaken interactions between the...SpCas9-derived variants with non-NGG PAM sequences include: xCas9 - NG, GAA, and GAT; increase nuclease fidelity...adeno-associated viruses (AAVs) . Other natural Cas orthologs include Hsp1Cas9 (from Helicobacter sp. MIT 11-5569 ) and...members inhibit CRISPR by a variety of mechanisms, including blocking crRNA loading or PAM recognition; inhibiting...

...Addgene's molecular biology reference guide, including information on molecular genetics, plasmids, sequencing...years, many significant discoveries lead to the conclusions that genes encode proteins and reside on chromosomes...essential DNA sequences for this purpose, which includes a DNA replication origin, an antibiotic-resistance...whose product confers resistance to ampicillin) is included in the plasmid. These bacteria are then grown ...in molecular biology for a variety of reasons, including that they are: Easy to work with - Plasmids are...gene expression in a wide variety of organisms, including plants, worms, mice, and even cultured human cells... a DNA fragment. Commonly used cloning vectors include Gateway entry vectors and TOPO cloning vectors....

...guide to using Optogenetics plasmids in your lab, including explanations of microbial opsin and optical switch... is a list of commonly used microbial opsins, including a brief overview of the wild-type opsin of each...functionality of ChR. Examples of ChRs from other species include: CsChR (from Chloromonas subdivisa ), CoChR (from...depolarization) of the neuron. Feature enhancements include: Increased photocurrent amplitude Examples: ChR2...Examples of anion channel variants from other species include: GtACR1 and GtACR2 (from the cryptophyte Guillardia...Guillardia theta ). Other feature enhancements include: Increased photocurrent amplitude Examples: iChloC,...Variants. Mac variants have been engineered to include enhancements such as: Improved photocurrent amplitude...

...content , including protocols, tips on using Addgene’s materials, and career advice. Playlists include: Protocols...Overview of Addgene's educational resources, including eBooks, science guides, videos, and protocols ...popular topics related to our collections. Titles include: Plasmids 101 CRISPR 101 Viral Vectors 101 Fluorescent...and related molecular biology techniques. Topics include: Genome Engineering Fluorescent Proteins CRISPR...assist with your molecular biology research. Topics include: Antibiotic Concentrations for Bacterial Selection...molecular biology, plasmid cloning, and virus, including: Gel Electrophoresis Bacterial Transformation ...

...2) to completely unrelated molecules. Examples include: scFv - Single-chain fragment variables (scFvs)... used to quantify relative amounts of protein. Includes: Western blot ELISA Capture - used to isolate ...isolate specific proteins or protein complexes. Includes: Immunoprecipitation (IP) Chromatin immunoprecipitation...Visualization - used to visualize protein location. Includes: Immunofluorescence (IF) Immunohistochemistry ...identify, analyze, capture, and sort single cells. Includes: Flow cytometry Read on to learn more about the...the applications common to each category, including an outline of the basic steps needed for each application...run. The standard curve should contain a range inclusive of all expected protein concentrates in the sample...

... may also be included within a transfer plasmid: for example, the U6 promoter is included in the pSico...expression. Other features that can be included in transfer plasmids include: Tet- or Cre-based regulation and...genome. Typically it is the sequences between and including the LTRs that is integrated into the host genome...ONLY by a second generation packaging system that includes TAT Can be packaged by both 2nd and 3rd generation...vector, some possible approaches to using this site include subcloning or appending compatible restriction ...

...controlled by various DNA regulatory sequences including enhancers, boundary elements, insulators, and ...of gene regulation, like operons. Operons are a cluster of different genes that are controlled by a single... shRNAs and gRNAs. Resources Addgene's blog , including our popular Plasmids 101 series , covers topics...

... target gene Many different activators exist, including the multi-plasmid SAM system. Select Expression...CRISPR in mammalian cells. Some of the variables include: Species of Cas enzyme and gRNA Species of promoter...verify that your cells contain your desired edit, including but not limited to: Mismatch-cleavage assay (for...

...involved in a wide variety of biological processes, including initiating signaling pathways in inflammation ... less than ideal. DREADDs Designer Receptors Exclusively Activated by Designer Drugs (DREADDs), like RASSLs... Relationship Studies for Designer Receptors Exclusively Activated by Designer Drugs. ACS Chem Neurosci...

...genome. Typically it is the sequences between and including the LTRs that is integrated into the host genome...internal promoter. An internal promoter must be included as the wildtype LTR promoter is no longer present...

...Variants As mentioned above, AAV improvements have included the production of synthetic capsids and the mixing...viruses with new properties. Other system variants include: Self-complementary AAV (scAAV): One downside of...

...of viruses that are commonly used in research, including: Lentivirus Adeno-associated Virus (AAV) Adenovirus...

...November 2018 | First Edition About The guide includes information on the many career options in science...

...selection mechanisms for lentiviral CRISPR libraries include: resistance to the anti-cancer drug vemurafenib...

...reference guide on molecular cloning techniques, including information on seven commonly used cloning methods...