...viruses that can infect people, cells, and animals. Some of these viruses are completely inert, but others ...protection are key to lab safety and the same principles apply to viral handling. Gloves are always required...

...others are studying and gives you a sense of what people are excited about. Following scientists in fields...more about authors by exploring author pages (for example, Melina Fan’s author page). These pages show all... Key features Setting up a personalized feed is simple. You can enter as many search terms as you like...

...poster should catch the audience’s eye and draw people in. That can be accomplished with an exciting title...clearly identified and interpreted in less than a couple of minutes. How does one accomplish all of that...column left-to-right, which is the natural way that people expect to read. The key to the success of this ...

...ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll find in this ... well-characterized Cas enzymes, which rely on multiple active sites. In addition, Casɸ has a minimal ...GFAP.SF-iGluSnFR.A184S-AAV1 here New Dual and Triple feature INTRSECT viral preps from the Deisseroth...

...anything. Viruses are complicated and they perform complex biological tasks, both of which are factors in ...Ugai et al., 2002). If the virus will be used in a couple of days, some researchers prefer to store freshly...: 16510845. 4. Krajden, Mel, et al. "Effect of multiple freeze-thaw cycles on hepatitis B virus DNA and...

...loxP, black arrowheads) and one mutated pair (for example- lox511, white arrowheads). It is necessary to ...express GeneR only in the presence of Cre. In this example, Cre would first invert the reporter and GeneR ...2008. A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range circuit ...

...modification is chromatin remodeling. Chromatin is a complex of DNA and histone proteins that compacts cellular...different disease phenotypes, are two fascinating examples of epigenetics in the form of imprinting. In imprinting...have huge impacts in the way we study and treat complex or multifactorial diseases, enabling faster diagnosis...

...RNA (pegRNA). The pegRNA consists of a target-complementary spacer, a scaffold sequence, and an extension...screens with oligo libraries where pegRNAs are coupled to their target allowing us to edit and evaluate...efficiencies for diverse prime editing systems in multiple cell types. Cell, 186, 1–17. https://doi.org/10.1016...

...AdV-mediated transduction of gRNA:Cas9 ribonucleoprotein complexes into transformed and non-transformed cells yielded...a potential platform that could be realized via simple switching of the relevant gRNAs and donor DNA Adenoviral...vector delivery of RNA-guided CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse array...

...with knockouts of specific genes. Many years ago, people in the laboratory became frustrated with using ...procedure (illustrated in the YouTube video below) is simple: After obtaining antibiotic-resistant colonies,...initially, or using secondary biological screens. For example, for cancer researchers, if growth properties in...

...Compared to humans and mice, flies have a relatively simple genome. They have four chromosomes: the X (and ...chromosomes essentially are messed up chromosomes with multiple inversions. They are unable to participate in ... dominant phenotypes visible on the flies. For example, second chromosome balancers often have the curly...

.... For more mechanistic details on HDR and NHEJ, please see the linked blog posts. Compared to its counterparts...strategy looks very similar to HDR, but it is completed using much shorter sequence homologies. Abbreviated...Genome-Editing Based Approach for Mapping Multiprotein Complexes in Human Cells.” Cell Rep. 13: 621-633. PMID: ...

...ribonucleoprotein (RNP), consisting of the Cas9 protein in complex with a targeting gRNA, to your cells of interest...CRISPR experiment is the generation of the RNP complex.  While one option is to purchase the Cas9 protein...bread wheat using CRISPR/Cas9 ribonucleoprotein complexes." Nature communications 8 (2017): 14261. PubMed...

.... Here is an example of a plasmid with a downloadable protocol listed under “Supplemental Document”: gRNA_Cloning...of checking off each step as you painstakingly complete your experiment – one small victory in a long ...

.... Now GFP and its fluorescent derivatives are a staple in the lab. GFP is used in research across a vast...importance of the entire GFP protein, which is almost completely required to maintain fluorescent activity; very...exist with only slight variations separating them.  Please note that many fluorescent proteins found on the...

...of DNA and both identified lymphocyte antigen 6 complex (LY6A) as a receptor for PHP.B. Read on to learn... the majority of astrocytes and neurons across multiple CNS regions. PHP.eB, an enhanced version of PHP.B... interested in using AAV-PHP.B vectors? See Supplementary Table 3 in Huang et al. for a list of PHP.B ...

...light inducible ion channels (plasmid # 20298 for example) have revolutionized Neuroscience. They are the...researcher exquisite control over expression of multiple genes in specific cells at specific times. Many.... “A FLEX Switch Targets Channelrhodopsin-2 to Multiple Cell Types for Imaging and Long-Range Circuit ...

...granularity, a proxy for nucleic complexity of the cell.  The sample will have been labeled using a panel...parallel.  Let’s imagine we’re looking for cells in a sample that are negative for marker A and positive for...

...polycistronic tRNA-gRNA (PTG) gene system to clone multiple protospacers which enables us to target more than...Minkenberg, and Yinong Yang. "Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing...PMC5972399. 12. Molla, K., Shih, J. & Yang, Y. Multiplex CRISPR-mediated base editing in rice. In Genome...

...system frequently over the past fifteen years. Some examples from their lab include using AAV to introduce ... alleles of a gene, or which require modifying multiple genes simultaneously in the same cell, are more...by using a targeted nuclease. Asuri et al, for example, used zinc fingers to increase the efficiency of...

...GCN4 binding scFv that’s fused to GFP. When the multiple copies of the scFv-GFP fusion bind the SunTag ..., Alitheen NBM, Hamid M (2012) scFv Antibody: Principles and Clinical Application. Clinical and Developmental....0000000000000145  Gąciarz A, Ruddock LW (2017) Complementarity determining regions and frameworks contribute...

...pluripotent stem cells (iPSCs). The CRISPR/Cas9 com­plex is very efficient at introducing double stranded...the PAM as possible (see below to find out why). Please check our recent studies for details (Paquet, Kwart... systems.     References 1. Cong, Le, et al. "Multiplex genome engineering using CRISPR/Cas systems." ...

...primary cellular receptor and sometimes one or multiple coreceptors, which may be present on different...and modifying capsid and genomic structures. For example, rAAV pseudotypes mix the genome of one AAV serotype...  AAV8 AAV8 has high transduction efficiency in multiple tissues, rapid transgene expression, and can cross...

... any gene of interest, one should require that multiple gRNAs of different sequences give rise to the ...but can often require difficult decisions. For example, would it be better to use a less-active gRNA that... in the interpretation of results by requiring multiple sequences targeting a gene to score in order to...

...allows for targeted delivery of Tet1 alone, or coupled with DNA oxidation and repair factors that allow...more frequently inherited by daughter cells. For example, KRAB induced silencing was transient and quickly...and context-dependent, it may make sense to try multiple CRISPR tools when setting up your experimental...

...extracting the DNA from fossilized amber and supplementing it with the DNA of present-day frogs via genetic...template.  Each zinc finger recognizes a nucleotide triplet and when several ZFNs are strung together, it can...the ZFNs, but instead of recognizing nucleotide triplets, the components recognize individual nucleotides...

...cassette (Dickinson et al., 2015) made this step as simple as heat-shocking to induce the expression of a ...direct injection of a CRISPR/Cas9 ribonucleoprotein complex with linear DNA as repair templates which reduces...worms are perfectly viable at 15 ºC, yet exhibit complete embryonic lethality at 25 ºC. This method resulted...

... viral DNA sequences and expression profile at multiple steps from plasmid amplification in bacteria to... set of injections into your animal model. For example, in a recent study by the Wickersham lab (Lavin...essential that every new vector, whether it be a completely new transgene or merely a new lot, be thoroughly...

...are working with proto-oncogenes or with human samples that have not been tested for HIV.  If not using...must be chosen carefully. Often researchers want a simple set-up where the gene of interest is expressed ...experience in transfection, CaCl2 transfection is a simple and cheap technique that should work well. Quickly...

...CRISPR-based gene editing (see table below). For example, the far-red light-activated split-Cas9 (FAST) ...translocation of the protein BldD to the nucleus. To couple this light-dependent activity to transcription,...proteins by fusing Cas9 to photoreceptors. One prime example was from the lab of Michael Lin, who developed ...

... Denise Roche, to edit the translation. We are pleased to announce that CRISPR 101: Una herramienta de...repository. This incredible gift will allow more people to access helpful resources for their CRISPR experiments...

...Created in collaboration with Addgene writers High Complexity Golden Gate Assembly with NEB's DADExperts: Matthew...KarplusIBEX Knowledgebase: A Data Resource for Multiplex Tissue ImagingResearchers: Andrea Radtke, Ziv ...

...pm. If you want to learn about, and practice, principles of clear, accessible writing through the lens...Poster Session TipsHow to Make Friends and Meet People at a Scientific Conference New to SfN? Try A Novice's...

... AAV performed in that experimental setup. For example, this is an experiment using the chemogenetic tool...thank you for your contribution to open science.  Please note that the AAV Data Hub is a beta project, and...

...was designed simply to be stable and novel, the principles that created it have been applied to numerous...protein evolution. The Baker Lab has deposited multiple Top7 plasmids at Addgene, along with other tools...

...preps. For example, last year we published an article describing a sucrose supplementation strategy to...

...Sleigh bells, please! It's time to announce the winners of the 2022 #DeckTheLab contest!  and it's.....I was bored this weekend while waiting to take samples from my assay at like 23:00, so I decided to make...

...Blog is a great way to get your name out there! Completed blog posts are also great to include on CVs and... with and discuss your work with many different people. Recent Guest Writers Have Included: Wenning Qin...

...basics - diving into the ins and outs, tackling complex questions, and giving tips for consistently generating... with some behind-the-scenes peaks of Addgene’s people, culture, and workplace. Rumor has it that a few...

... (AAVrg), and volumes of 1 mL, 2.5 mL, or 5 mL. Please note that all orders will be shipped in 100 µL ...are a big deal, so if you have questions, we have plenty of information available for you to dig into. Check...

..., in vitro screens cannot faithfully model the complex spatial, cellular, metabolic, and cytokine microenvironment... cells, an important immune cell type, in more complex and physiological microenvironments. These screens...

...just starting in the lab, but they are exactly the people you should talk to if ruminating the trials and... 14:32 - 16:07 : What do you think drives other people to do science? 16:08 - 22:04 : How have the make-up...

...know-how to transcend national borders and help people around the world. As part of our own mission, we...Mentorship Program Learn How to Network and Meet People at Scientific Conferences Resources on Addgene.org...

...cultural shifts that can have bigger impacts. For example, if you joined an office where everyone used a ... about this, and even has a list of biological samples that are stored long term at -70 °C. My Green Lab...

...contributor of careers blog articles, Joanne had plenty of advice for the group of juniors and seniors,...working lab full of busy Addgenies preparing plasmid samples for shipping. This was the favorite part of their...

... in the wired world we are living in, there are plenty of other ways to define and measure scientific ...measure will ever adequately capture a notion as complex as “scientific impact.” As we look ahead to the...

... While those are the biggest changes, there are plenty of less obvious ones to discover as you browse.... Multiple Authors...

...experiments. Finally, we were thrilled by how many people accessed our freely available online collections...the promise of science to help a great number of people. I look forward to working with Addgene colleagues...

...allow the extraction of enough plasmid to last for multiple experiments. Another fundamental aspect for transfection...nucleic acid and chemical contaminants, these DNA samples must also be entirely endotoxin free. Endotoxin...

... which display sophisticated interactions with multiple other proteins. Again, little to no change in ... place to start. These tags are easy to use in multiple common applications. Due to their limited effects...