We narrowed to 883 results for: prot

...desired edit (or edits) engages the prime editor protein. The prime editor consists of a Cas9 nickase fused...they engineered a dominant-negative mutant of the protein MLH1, a component of the MutSα–MutLα mismatch repair...strategies to evolve more effective prime editor proteins, but found that the mutations that emerged depended...with improved stability Traditional sgRNAs are protected from cellular degradation by Cas9 binding, but...target site, but it cannot drive prime editing. To protect the 3′ end, the Liu lab appended RNA pseudoknots...stability. Using a CRISPRi screen, they identified a protein that enhances prime editing in mammalian cells:...cells: the small RNA-binding exonuclease protection factor La. They found that La, which is ubiquitously expressed...

...periplasmic glutamate binding protein known as GltI. When the resulting fusion protein binds to glutamate, conformational...Choosing the B(right)est Fluorescent Protein Visit the Fluorescent Protein Featured Topic Page Resources at...Addgene.org Find Fluorescent Protein Biosensors Browse All Fluorescent Proteins Find AAV Tools References...vector, use with IPTG induction to express soluble protein CAG: Generic strong promoter. Formulated as AAV-DJ...FLP-dependent expression GFAP: Glial fibrillary acidic protein (promoter). Good for Glial expression (not neurons...

.... Once a co-repressor protein binds the repressor protein, the repressor protein can bind to the operator...activator protein is bound, turning transcription ON. When a repressor binds the activator protein, the activator... and largely defines the amount of recombinant protein that will be produced. Many commonly-used promoters... activator protein cannot bind the promoter sequence anymore and transcription is turned OFF. Negative...Negative Repressible: A repressor protein is present, but cannot bind the promoter sequence and transcription...expression system for sphingomonads and other Alphaproteobacteria. Appl Environ Microbiol. 2013 Nov;79(21):...

...vesicular somatitis virus G glycoprotein (VSVG). VSVG is a broad tropism envelope protein that can also be pseudotyped... at first to look at so many plasmid maps and protocols! In reality, there are many shared components ...interact with the host. This is the role of Gag proteins: they contain the matrix, capsid, and nucleocapsid... Viral packaging and entry The viral packaging proteins for AAV are rep and cap. Rep is required for viral...AAVS1 locus in humans. Cap is a structural capsid protein that helps with virion assembly and particle release...discussed. LXs/Iva2 are primarily capsid assembly proteins required for packaging of the virus into particles...Resources Kulanayake, S., Tikoo, S. K. Adenovirus Core Proteins: Structure and Function. Viruses. 2021 13(3):388...

... accurately run a protocol.   Figure 1: Evolution of a Western blot protocol, from a full page ...checklist for everything you do in the form of a protocol, and you can add in personalized steps like “check...re-sterilize now.” Have a physical copy of the protocol when you work and make a checkmark next to each... it anyway.  You may find that after running a protocol a few times, you no longer need the full checklist...check things off every 3 steps, or you rewrite the protocol and consolidate several steps into one. Soon you...as you run your experiments. Try drawing out a protocol as you read it, or putting a diagram up on your...may seem counterintuitive to add more time to a protocol if you’re in a hurry, but let’s consider it this...

...CRISPR-Cas9 system. Nature Protocols, 8(11), 2281–2308. https://doi.org/10.1038/nprot.2013.143 Sander, J.... compared to where the target site sequence (protospacer+PAM) is located? If there are other PAMs in the... there needs to be extensive homology of the protospacer followed by PAM. See Addgene's CRISPR resources.... In the case where you have intact CRISPR protospacer target in the HR template, you usually would ...make sure it's a silent mutation). Q8: Is a protospacer followed by the PAM (my target sequence) in my...template an issue? A8: Yes. If you have intact protospacer+PAM sequence within the HR template, it will ...addition to mutations in the spacer itself, will protect the HR template from the Cas9. Again, if it's within...

...new Cas enzymes on the block? The Cas family of proteins has many useful genome engineering enzymes, each...engineering purposes far more than any other Cas family protein and has thus benefitted from countless rounds of... RNA editing. Fast facts – PAM requirement: protospacer flanking sequence – A, U, or C. Best for: targeting...development and use. Fast facts – PAM requirement: protospacer flanking sequence – none. Best for: targeting...   Table 1: Fast facts on Cas family proteins and genome editing abilities   Choosing a...adaptable Cas currently available, the other Cas proteins should not be overlooked. The unique features ...al., Programmable RNA targeting with the single-protein CRISPR effector Cas7-11. Nature, 597, 720-725 (...

... the proteins the transposon needs for transposition. Transposition protein: This is the protein that’...sequence to move from one DNA site to another. This protein is either a transposase or a reverse transcriptase...They contain two genes: gag and pol. The pol polyprotein encodes the reverse transcriptase and integrase...as “complete transposons” since they encode the protein they need to move around, either a transposase ...major components: 1) a CRISPR RNA (crRNA), 2) four proteins (TniQ, Cas8, Cas7, Cas6) that forms the QCascade...transposase complex made up by three transposase proteins (TnsA, TnsB, TnsC), and 4) the donor DNA or mini-transposon...transposon’s DNA sequence and its transposition protein, to move DNA. There are three popular systems for...

...Genetic Engineering. Nature protocols. 2009;4(2):206-223. doi:10.1038/nprot.2008.227. PubMed PMID: 19180090...other small base pair changes, and the addition of protein tags. It also has the flexibility to modify the... complementary strand was degraded. Beta: Beta protects the ssDNA created by Exo and promotes its annealing...: 1) use a strain of bacteria that has key MMR proteins knocked out or 2) specially design ssDNA oligos... not the amino acid sequence of the translated protein.  These changes can be 5’ or 3’ of the desired ... detail below).  Controlling expression of Red proteins is critical for minimizing the toxic effects of...Bacteria Using Homologous Recombination. Current Protocols in Molecular Biology. 106:V:1.16:1.16.1–1.16.39...

...existed! Addgene recently started automating a few protocols to make standard lab processes easier and more...own lab. Automating bacterial transformation One protocol that has been automated is high throughput transformation... machine that would improve our transformation protocol and, after looking at a few options, we decided... to use the FXP to automate and optimize more protocols in the future - something that will be key for...same amount of time by hand! Setting up a basic protocol on the Biomek FxP is pretty simple. By clicking...perfectly. Working with Qiagen, we developed a protocol to run up to 192 samples at a time. It follows...contamination between samples. With this automated protocol, we can prep two 96 well plates in just under ...

... of neural circuits. Nature protocols. 2012;7(1):12-23. doi: 10.1038/nprot.2011.413. PubMed PMID: 22157972..., most of which is relatively cheap). Several protocols have been written outlining the different steps...steps and materials required for each [1, 2]. This protocol [3] provides, in exquisite detail, the various...ferrule implant must be maintained during the entire protocol, otherwise optical stimulation will be erratic...generation (TTL pulse) is a standard electronics protocol not specific to lasers or optogenetics so cheaper...Optogenetic control of motor coordination by Gi/o protein-coupled vertebrate rhodopsin in cerebellar Purkinje...channelrhodopsin for activation of CNS neurons. Current protocols in neuroscience / editorial board, Jacqueline ...

...-F, but instead employ a Cas12k protein instead of three Cas proteins (Fig. 1A, bottom), and require only...spacer sequence specifying the target site. Four proteins (TniQ, Cas8, Cas7, Cas6) that forms the QCascade...module together with the crRNA.  Three transposase proteins (TnsA, TnsB, TnsC). The donor DNA (aka mini-transposon...INTEGRATE. The requirement for multiple Cas proteins to form the Cascade targeting complex is a hallmark...Type I CRISPR-Cas systems, in contrast to single-protein effectors employed by Type II systems (such as ...forms a novel co-complex with the transposition protein TniQ for targeting (Halpin-Healy et al., 2020).... only two transposase proteins (TnsB and C). While requiring fewer components for RNA-guided DNA integration...

...well-established protocols for their production and purification that yield high titers (see also AAV protocols for...Maday et al., 2014). It is currently unclear which proteins facilitate transport of neuronal tracing compounds... For predominantly retrograde tracing, albumin protein labelled with horseradish peroxidase (HRP; Kristensson...contained by a capsid which is comprised of the viral proteins VP1, VP2, and VP3. To date several hundred variants...engineered to have differences in their capsid proteins. For a brief overview see Addgene’s AAV guide....neuron-specific transgene expression. Fluorescent proteins are widely used for tracing and their expression...that express one of a few different fluorescent proteins (red, green, blue). If a researcher injects a ...

...tagging Lentiviral capsid-based Cas9/gRNA ribonucleoprotein delivery Cytosine deaminase for RNA editing... Burkholderia, they identified 67 genera of Proteobacteria that this toolkit might be used in. Read the...engineered SECURE adenine base editor variants using a protein-truncation strategy with reduced RNA-recognition...abilities of C-to-T and A-to-G base editors into one protein. They fused an activation-induced cytidine deaminase...bioRxiv Find the ORANGE toolkit   Cas9/gRNA ribonucleoprotein delivery by lentiviral capsid-based bionanoparticle...bionanoparticle system for packaging Cas9/gRNA ribonucleoproteins for transient expression. They replaced the...with a com aptamer which binds an aptamer binding protein for packaging into a lentivirus-like particle. ...

...permuted green fluorescent protein (GFP), and nLightR uses a red fluorescent protein, mApple. Kagiampaki et...that red fluorescent proteins tend to be less sensitive than green fluorescent proteins. nLightR did perform...S., & Steinegger, M. (2022). ColabFold: Making protein folding accessible to all. Nature Methods, 19(6...More resources on the Addgene blog Fluorescent proteins 101: Fluorescent Biosensors Which Fluorescent ...Find Optogenetics Plasmids Visit the Fluorescent Protein Guide Pages ...

...setting. Read more: Plasmids 101: Protein Tags Produce proteins that glow so that scientists can track... a plasmid by recruiting replication machinery proteins Antibiotic Resistance Gene Allows for selection...gene is expressed in and amount of recombinant protein obtained. Selectable Marker The antibiotic resistance...used to do include: Produce large amounts of a protein so that scientists can purify and study it in a...inside a cell Plasmids 101: Green Fluorescent Protein (GFP) Plasmids 101: Luciferase Monitor the ...plasmid stocks. The guide also contains multiple protocols and troubleshooting tips to make plasmid usage...

...recombinant proteins at high levels. In a case where clinical applications of recombinant proteins are necessary...to the production of clinical viral therapies. Protein production  HEKs are extremely easy to transfect...eliminate non-human PTMs which is a concern with other protein production systems, such as yeast. HEKs can also...cell line’s ability to faithfully produce these proteins and its relatively low expression of native channels...receptor biology, ion channels, or involve human protein preparations with PTMs. Why the HEK not? References...Cell Line as a Platform to Produce Recombinant Proteins and Viral Vectors. Front Bioeng Biotechnol. (2021...

...fluorescent protein followed by a stop codon and the coding sequence for a green fluorescent protein (GFP)....biochemical secondary energy sources to increase protein yields from 36 μg/mL to 266 μg/mL. The reaction...-free system, capable of producing fluorescent proteins and enzymes. Image from Moore et al., 2021. ...through monitoring the expression of fluorescent proteins and oxytetracycline enzymes. Next, this system...libraries: Cas13X and Cas13Y, two compact Cas13 proteins found in the metagenome datasets from hypersaline...messenger-RBP sgRNA Library targets human mRNA binding proteins with 10 guide RNAs per gene. The lab modified ...

...concentrating your virus. See our Concentrating AAV Protocol (Step 10) for instructions on how to do this. ...necessary to visualize expression of fluorescent proteins, for example, as many viral particles must infect...infect a cell to achieve visible expression of the protein. However, expression of receptors may not need ...In the case of channelrhodopsins, fewer channel proteins are required for the transduced cell to become...become light responsive than fluorescent proteins required to get a good fluorescent signal. It may not be...take below but you should read our AAV Production Protocol for additional best practices. Keep your cells...Addgene.org Find Ready-to-Use AAV Preps Find Lab Protocols for Work with Virus Watch Our AAV Purification...

...structural component of atherogenic lipoproteins, shuttles lipoproteins through the circulatory systems, ...LipoGlo: A reporter for measuring ApoB bound lipoproteins Article contributed by Leah Schwiesow A leading...cardiovascular disease is driven by atherogenic lipoproteins that accumulate within the vascular wall. ApoB...formation of atherosclerotic plaques. ApoB bound lipoproteins (ApoB-LPs) are indirectly measured through their...requires binding of bacterial enhancer binding proteins (bEBPs) at a sequence upstream of the promoter...fused to an RNA binding motif, and a RNA-binding protein-bEFP fusion. Thus, the bEFPs bind the UAS via the...