...add a bit of fun. He mixed together 10 E. coli strains producing different pigments, and tasked the students...perfect fit for the streak plate challenge. Chromoproteins are a subset of the fluorescent protein family...off color in ambient light. Expressing these chromoproteins in E. coli would make it easy to distinguish...single colonies, these chromoprotein-expressing strains are easily amenable to other experiments to teach...have the students transform them into different strains of E. coli,” says Westenberg. During the transformation... how different genetic backgrounds of the host strains affect transformation efficiency.  Aside from the...blog Read our other blog posts on fluorescent proteins Check out educational resources on the blog Resources...

...coding genes in the human genome with fluorescent proteins. For each gene, we needed to design guide RNAs...ranked by specificity of targeting or by cut-to-insert distance for HDR. (The notable exception is TagIn...compressed CFD score) * (gaussian normalized cut-to-insert distance) This gives each guide a score ranging... can be used in the rank equation above. Cut-to-insert distance was fitted to experimental data of its...the greater this distance between the cut and the insert, the more poorly the guide performs HDR. Optimal...sequences to prevent re-cutting of the sequence inserted by HDR. Although this practice is generally advised...enough to prevent re-binding. Lastly, the sequence inserted by HDR may inadvertently re-create a match for...

...modulation of endogenously and exogenously expressed proteins. Here we will review what advantages degron tags... the tagged protein. Degrons are destabilizing domains which will be identified by various enzymes and...manipulate endogenously and exogenously expressed proteins. Thirdly, degrons and their drugs are titratable...not amenable to being tagged, e.g. tagging some proteins can affect their function, the SMASh tag self-...developed as an affinity tag for visualization of proteins of interest. A fluorescent Halo ligand can be ... molecules which target E3 ligases to specific proteins by simultaneously binding both their target protein...their effectiveness with over a dozen different proteins. In the end, you can’t predict with absolute certainty...

... pC5Kan-P2A P2A Insect pUltra P2A and T2A Lentiviral Ac5-STABLE2-neo T2A Insect   For the co-expression...vectors simultaneously express two or more separate proteins from the same mRNA. We've discussed promoters ... detect your gene of interest, scientists have instead developed novel methods to co-express your gene...IRES Elements Translation in eukaryotes usually begins at the 5’ cap so that only a single translation...site, thereby resulting in co-expression of two proteins from a single mRNA. IRES was originally discovered...between your genes of interest and you can then insert the whole multicistronic cassette into a backbone...Although it is recommended to use the 2A peptides instead of an IRES when stoichiometrically equivalent levels...

...translation of RNA to make proteins. That's central to everything that goes on inside a cell. Until we get ...of enough sensor domains. Ultimately, we'll need a couple thousand or so sensor domains for everything ...DNA systems capable of storing data and computing inside living cells. He's also a co-founder of the BioBricks...BioBricks The BioBrick Foundation's Linda Kahl explains the Addgene collaboration this way: "Addgene is...strategy that organizes how we start the work, and it begins with the central dogma - replication of DNA, transcription...and expensive. There's the promise that we could instead have things like peptide or RNA aptamers that act...

...for Science.  Cold and overcast with intermittent rains, it was weather befitting of the ominous political...Science and Engineering at Rensselaer Polytechnic Institute. He also studies energy markets in his spare time...cause breast cancer and infertility or to argue against unnecessary clinic regulations, evidence-based ...issues hit hardest. According to the Guttmacher Institute, most women of reproductive age live in a state... how planets form around young stars and the instruments we use to study them. You can follow her at alexandrazgreenbaum.wordpress.com.... I worried that, because we were speaking out against legislators and because the march had been so well-publicized...scientists – including global warming skeptics – insisted that more data and research funding are critical...

...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert Promoter PI...Drosophila ID Plasmid Gene/Insert Promoter PI Publication Plant ID Plasmid Gene/Insert Promoter Selectable Marker...elegans ID Plasmid Gene/Insert Promoter PI Publication Yeast ID Plasmid Gene/Insert Promoter Selectable Marker...

...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert Promoter Selectable...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Plant ID Plasmid Gene/Insert Promoter Selectable...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type...PI Publication Yeast ID Plasmid Description Gene/Insert Promoter Selectable Marker PI Publication Do you...

...Harrach, 2014). Given that the early genes encode proteins essential to initiating the whole process – such...used to house a recombinant sequence (transgene) instead.    Although the late genes are usually present...absent, and the late genes (and their structural proteins, which would encapsidate new virions) are not ... is transient due to the fact that AdV surface proteins (especially fiber’s knob domain, and hexon), as... as well as other intracellularly-exposed proteins and downstream cellular events, all recruit strong ...sense of what AdVs are about – and we hope that it inspires further use of existing vectors, as well as research...resources References Bauzon, M., Castro, D., Karr, M., Hawkins, L. K., & Hermiston, T. W. (2003). Multigene expression...

...now available. In this post, we’ll review the inspiration behind ExM, how it works, and some of the limitations...most SRM techniques also require specialized instruments and technical expertise, limiting their accessibility...a straightforward answer to the space problem. Instead of imaging smaller, what if we just made the sample...chemical dyes, and genetically encoded fluorescent proteins (Gambarotto, et al. 2019; Tillberg, et al. 2016...adapted for detecting nucleic acids as well as proteins (Klimas, et al. 2023; Wassie, et al. 2019). Perhaps...et al (2021) Expansion microscopy provides new insights into the cytoskeleton of malaria parasites including...Saad O, Bewersdorf J (2020) Light microscopy of proteins in their ultrastructural context. Nat Commun 11...

...classified as a type V CRISPR system. Like Cas9, Cpf1 contains a RuvC-like endonuclease domain, but it lacks ...off-target editing efficiency. Kim et al. and Kleinstiver et al. characterized genome-wide editing efficiency... to single mismatches, but variably so, with Kleinstiver et al. reporting that Cpf1 can tolerate mismatches...essential function in Cpf1-mediated editing, as Kleinstiver et al. saw no decrease in Cpf1 activity with ... are indicated below each graph. Figure from Kleinstiver et al., 2016. Cpf1’s strength may lie in ...activity than that previously observed with SpCas9. Kleinstiver et al. suggest that AsCpf1’s off-target rate ...eSpCas9 and SpCas9-HF1. Both AsCpf1 and LbCpf1 ribonucleoproteins (RNPs) failed to induce off-target editing...

...to learn about the applications of fluorescent proteins and to find the plasmids that are available from...Monomer not a Monomer? Addgene's plasmid repository contains a variety of fluorescent protein plasmids. Use...more about the many applications of fluorescent proteins (FPs) and to find the plasmids that are available.... Regulate Biological Activity Use fluorescent proteins to modulate biological activity, like transcription...consisting of ORFs tagged with a variety of fluorescent proteins, as well as over 100 empty backbones. Fluorescent...Jin Zhang Lab at UCSD Properties of Fluorescent Proteins from Kurt Thorn and UCSF Molecular Expressions...

...days inside the mother, the zebrafish is an ideal system to observe embryogenesis live. The origins, divisions...implemented in zebrafish. Developmental biology insights using lineage tracing in zebrafish To highlight...lineage tracing approaches that led to important insights in developmental biology. Microscopy-based lineage...example, neuronal activity can be tracked in whole brains of swimming, actively learning larvae, providing...resistance (so far) to facile CRISPR/Cas9-mediated insertion of foreign DNA. Despite these caveats, several...highlight how these advantages can translate into insights in developmental biology, and advances in this...zebrafish behavioral studies Download the fluorescent proteins 101 eBook Read our plasmids 101 blog series Resources...

...of the viral capsid proteins are analyzed. The abundance of viral capsid proteins as a fraction of total...relative stoichiometric ratios of the viral capsid proteins VP1, VP2, and VP3. Samples of viral preparations...subjected to gel electrophoresis. Viral capsid proteins VP1, VP2, and VP3 are shown relative to the total.... Three to five days later, cell cultures are inspected for sterility. Transducibility Some viral vectors...virus from Addgene, read our detailed recipient instructions for next steps. Plan an experiment using our...plasmid in the viral preparation by performing PCR against unique regions of the transfer plasmid.   Lentivirus...

... In this iteration, the size of the lentiviral insert was minimized to reduce recombination and avoid...transcriptase enzymes, inhibition of host cell proteins, and modifications to the pegRNA. The Britt Adamson...nucelocapsid protein, one of the virus’s most abundant proteins and a critical component for viral genome packaging...-LV [+M], negative control) were immunoblotted against Anti-SARS-CoV-2 Nucleocapsid Protein antibodies...middle panel). Samples were also immunoblotted against vinculin as a loading control (right panel). Image... of ssAAV, a targeting sequence of miR124T was inserted into the vector to silence transgene expression...editing, and delivery of disease-relevant large proteins. They demonstrated REVeRT in various contexts ...

...form that can be understood by others, balanced against how often such data is likely to be accessed. Adding...not averse to making data available, but there remains concern about the extra work this may entail.  ...parsed by someone other than the actual author. Instead, selected images are said to be representative ...another issue of proprietary data formats. Many instruments store raw data in formats that require specific...supported/readable. Is the author or their host institution expected to provide data in formats exported ...integrate datasets with their resultant figures. Instead of providing a “dead end” figure or table, clicking...natural, albeit unintentional, tendency to work against transparency. This opens science to charges of ...

...Neuroscience : Antibodies targeting proteins for neuroscience research. Institute for Protein Innovation Collection...other types of affinity reagents such as scFvs, DARPins, Fabs, and secondary antibodies. To learn more ...Collection : Antibodies to highly conserved proteins and other difficult-to-target antigens that are intractable... represent the official views of the National Institutes of Health. Need Some Help? Browse some of our... Antibody Guide Antibody Protocols Recipient Instructions Antibody Blog Posts Addgene Help Center...

... libraries. Find tools for CRISPR, fluorescent proteins, optogenetics, biosensors, and more....Addgene Plasmid Collections Addgene's repository contains 156,627 plasmids . We've gathered our curated ...Cre-lox TALENs Zinc Fingers Luminescence Fluorescent Proteins Luciferase Optogenetics Chemogenetics Viral Plasmids...Click here to view all kits. Kit Types Fluorescent Proteins Cloning Genome Engineering Pathways & ORFs Synthetic... Depositor Collection Types CRISPR Fluorescent Proteins Protein Expression Synthetic Biology Viral Vectors...RESOLUTE Plasmid Collection for Solute Carrier Proteins (SLCs) Structural Genomics Consortium Zinc Finger...

...cleaves precursor proteins to a biologically active state. The SARS-CoV-2 S protein contains a potential cleavage...credit: Maya Kostman for the Innovative Genomics Institute Addgene COVID-19 Resources The following types...distribute SARS-CoV-2 plasmids. The collection contains: Plasmids for E. coli and Yeast expression Spike...Return to top Plasmids Encoding Mammalian Genes or Inserts Several mammalian genes have been identified as...SHERLOCK and DETECTR . Feng Zhang's lab at the Broad Institute has utilized the natural RNAse activity of the...UnLOCKING (SHERLOCK and SHERLOCKv2). The Broad Institute has published information and protocols for utilizing...

...that red fluorescent proteins tend to be less sensitive than green fluorescent proteins. nLightR did perform... corresponding pixel-wise ΔF/F0 heatmaps. White insets indicate surface expression of the indicators over...based on alpha-1a adrenergic receptor; nLightG contains circularly permuted green fluorescent protein ... alpha2-AR antagonist, nLight sensors are not. Instead, they are blocked by alpha1-AR antagonist. This...and resources References Feng, J., Zhang, C., Lischinsky, J. E., Jing, M., Zhou, J., Wang, H., Zhang, ...More resources on the Addgene blog Fluorescent proteins 101: Fluorescent Biosensors Which Fluorescent ...