...looked hard at existing tools. As has been written here on Addgene, there has been a Cambrian-like explosion... for the CZ Biohub project by Manuel Leonetti to tag all 22,000 protein coding genes in the human genome...high-throughput design and analysis of CRISPR experiments. It has certainly boosted our productivity in the Biohub.... tools which were optimized for HDR, except for TagIn. HDR requires the additional step of mutating guide...cut-to-insert distance for HDR. (The notable exception is TagIn––see their "summarisation score".) For CrispyCrunch...

...insert FLAG tags into the genome of the pathogenic fungus Aspergillus fumigatus, which has been difficult...sequence between the microhomologies.MMEJ has certain advantages over HDR. Some species don’t have good ...vectors. Addgene depositor Takashi Yamamoto’s lab has harnessed MMEJ to create a new method for CRISPR ...of press. However, this process accounts for a percentage of the mutations seen with TALENs and CRISPR....follows. For increased versatility, it would be advantageous to adapt PITCh to insert genes into AAVS1, the...using the nuclease Cpf1. Since Cpf1 cuts in a staggered pattern, it is thought to be ideal for HDR-independent...and Ling Lu. (2016). “Highly efficient CRISPR mutagenesis by microhomology-mediated end joining in Aspergillus...

...also the fastest test that has been developed so far.  A similar method has been optimized to detect the...Two-step-SHERLOCK has a sensitivity in the zeptomolar range while One-step-SHERLOCK has a sensitivity in...using lateral flow strips. Since the probes are tagged with streptavidin, the lateral flow strip first...State University. Since its appearance, SARS-CoV-2 has spread to almost every part of the world manifesting...full-fledged pandemic. Containing the spread of this virus has become an utmost priority for countries around the...the outbreak ultimately flattening the curve, it has become evident that widespread testing is crucial... 83 known COVID-positive samples tested, DETECTR has a positive prediction rate of 95% and a 100% negative...

...to allow in-frame deletions and introduce epitope tags and RNA transcription start sites. The lab further...packaging, but their activity in mammalian systems has been in question. Three different labs have recently...CRISPR-Cas system for mammalian use Stanley Qi’s lab has developed a miniature Cas system (CasMINI) engineered...

...until the pups are 2-4 weeks old and use the ear tag method (creating different patterns of ear holes ...Samantha Young. The use of CRISPR/Cas9 for gene editing has expanded since its adaptation for use in mammalian...biology surrounding a particular gene (although it has been quite successfully adapted for use in screening... method for validating your selected gRNA’s, and has the bonus benefit of supplying you with primers for...sturdier than the delicate RNA and therefore there has less chance of contamination with RNase leading to...products will give specific details on what exactly has been inserted or deleted. Each lab will use a slightly... et al. "Feasibility for a large scale mouse mutagenesis by injecting CRISPR/Cas plasmid into zygotes....

...FPs) offer scientists a simple yet powerful way to tag cellular proteins and investigate protein localization...fusion, etc). For example, by engineering PA-GFP tagged to a mitochondrial matrix protein (mito-PAGFP) ...Markus, and Wei‐Lih Lee. "Photoactivatable GFP tagging cassettes for protein‐tracking studies in the budding...high contrast ratio usually indicates that a PA-FP has a low level of spontaneous fluorescence in the absence...an FP-chimeric protein may be at any one of many stages of synthesis and degradation. For this reason it...PA-FPs by performing several rounds of random mutagenesis screening for enhanced RFP variants. PA-mCherry...negative” photoswitching. In 2008, scientists used mutagenesis screens to find a version of Dronpa that doesn't...

... applications (v82 has higher affinity for glutamate but slower kinetics; v857 has faster kinetics but...but lower affinity) and with a variety of tags for localization (PDGFR, GPI, and SGZ). Read more from the...iGluSnFR3 (Aggarwal et al. 2022). This new version has higher signal to noise ratio and is better localized...for two-photon microscopy. The resulting JEDI-2P has faster response kinetics and increased brightness...and deep-tissue imaging. Vladislav Verkhusha’s lab has generated a new genetically encoded calcium indicator...interest, it’s important to choose a biosensor that has a binding affinity close to the range of concentrations...physiology and metabolism under various conditions. LiLac has improved response size and reduced sensitivity to...

...epitope-targeting antibody that is conjugated to a biotin tag but not a fluorophore. In a separate step, cells ...interest. Indirect labeling is achieved through protein-tag or protein-protein interactions, like biotin-streptavidin...selecting the primary antibody host species, as it has to be different from the other antibodies in your...target marker, allowing for its detection. The advantage of this system is that streptavidin conjugates...the primary antibody (Figure 1b). As above, the advantage of this system is increased flexibility for panel...

...localization 1 TagBFP 399nm/ 456nm histone 2B (H2B) Chromatin 2 Cerulean 433nm/ 475nm Lyn-tag (derived from...the newly generated MCS. MXS-chaining also has the advantage of being directional, meaning that (in the... cloning. MXS Chaining is one such technique and has been used to produce complex fluorescent reporter...culture systems (their original goal). Their work has demonstrated that the MXS-chaining method can be ...

...Post About Yeast Vectors Read our Blog Post About Tagging Yeast Genes ...cells where binding between the proteins of interest has occurred. Although the original Y2H systems utilized... number of different bait and prey vectors. This has been shown to be as effective as using five independent...transmembrane proteins, a split ubiquitin system has been devised (8). References 1. Casadaban M.J.,...activator (1a) or a repressor (1b). Scientists took advantage of these discoveries to create a new system for...subset based on a cellular compartment or growth stage). A prey library can either be applied to the bait...

...the the developing zebrafish pancreas. They first tagged these cells using plasmids. Then using HOTcre, ...more commonly known: the fruit fly. The fruit fly has been used in scientific research for over a century... a transgenic mouse line. Over the years the fly has become an ideal model organism to study an array ...development, genetics, and the nervous system. The fly has also emerged as an important human disease model ... described in 1993 by Norbert Perrimon’s lab and has been continuously improved upon since then. If you...Zebrafish (Danio rerio) Since the 1960s, the zebrafish has gradually become an important model organism. They...Caenorhabditis elegans) The worm, Caenorhabditis elegans, has been used extensively as a model organism since the...

...received a whole set of cards (Promoters, Inserts, Tags, Origins of Replication, Resistance gene and Selection... As I wrote in the blog post, BUILD YOUR PLASMID has been developed to teach gamma-retrovirus or lentivirus-based...

...any real traction.  This can carry a heavy price tag, especially if you're working on something other ...this deter your ambition. If you believe your idea has potential to make a difference, then make sure you...Boston-based startup focused on empowering early-stage scientists. You can think of miLab as a sort of ...biotechnology investment fund that would apply a percentage of returns to support student projects, but it...for such a fund is an impossibility at our early stage.  After each pitch we made, we considered the feedback...against you.  As I mentioned, you start at a disadvantage in the health science space if you don’t have...

... cloning procedure adds both V5 and His6 epitope tags to the C-termini of Met proteins, which can be used...relatively confident that the gene from the test species has a similar function to the gene in the reference species...left with Lab Director Doug Warner on the right) has taught courses in biochemistry, molecular cell biology...

...wrmScarlet11 which is less than 60 nt in length to tag the target protein. The team validated this tool ...generated with mRNA display (FingRs), Xue Han’s lab has developed AAV vectors to selectively visualize excitatory...engineered Streptococcus canis Cas9 nuclease that has a broad 5′-NNG-3′ PAM. Get the plasmids at Addgene... human and plant cells. Its small size offers advantages for cellular delivery. Get the plasmids at Addgene...

... Choose the AAV capsid or lentiviral prep needed Tags: Identify a specific marker protein Use: Similar...useful in an experiment, and want to see if someone has already created and deposited it with Addgene. Combining...resistance gene(s) used for selection in bacteria. Has Viral Service?: Allows you to filter results to only...

...et al. "AAV-mediated anterograde transsynaptic tagging: mapping corticocollicular input-defined neural...In order for an AAV particle to infect a cell, it has to come into physical contact with that cell. Since...

...Addgene is certainly going to have to find ways to tag and curate reagents from these emerging organisms...RNA and can express fluorescent proteins. This lab has created an automated tank for feeding and maintenance...scolopes (Joshua Rosenthal) The Hawaiian bobtail squid has an extreme number of nerve cells and an unusually... injection (hard to do, expensive, not every lab has the capability) with receptor mediated endocytosis...

...specific edit, such as the insertion of a fluorescent tag or the introduction of a specific mutation, one generally... Broad Institute, John Doench. CRISPR technology has made it easier than ever both to engineer specific...databases annotate the TSS in different ways, and it has been shown that the FANTOM database, which relies...mismatches that can nevertheless still lead to activity has been established, and can be used to identify sites...Perturbation Platform at the Broad Institute and has worked with many Addgenies to help improve the understanding...e141–e141 . https://doi.org/10.1093/nar/gkw583 Rees HA, Liu DR (2018) Base editing: precision chemistry ...

...studies when used as part of a two part fluorescent tag-then-express system that allows for controlled induction...bloodstream allows for dispersion of the virus but it has to be able to cross the blood-brain barrier to reach...treatments for neurological diseases). The Gradinaru Lab has a lot going on. Here are the key takeaways: Two...