We narrowed to 695 results for: prot

...PS: they’re free. 3. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection by Joachim...strategies for generating GFP fusion proteins. From considering protein size and shape, linker sequences,...optogenetics, chemogenetics, plasmids, fluorescent proteins, careers, and more. These resources can now be...Fluorescent Voltage Indicators, and Photoswitchable Proteins by Cary Valley, Shreya Vedantam, Michelle Cronin...used in cloning. 8. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi by Jennifer Tsang...and find plasmids containing these fluorescent proteins at Addgene. 7. Designing Your Chalk Talk for ...

...3: Share protocols and spreadsheets with lab members Now that you’ve organized your protocols, wouldn’... and your lab? Organizing your lab notebooks, protocols, and to do lists can be easier than you think!..., and post-its to make finding your most used protocols & references easy to find It’s happened to all...can’t remember where you put your flow cytometry protocol. Or maybe you can’t find the molecular cloning... visibly labeled on the spine. For binders of protocols and data, consider using color coded binders and...your colleagues. Start small by identifying the protocols, guides, or references that your labmates ask ...history. Now you’ll be able to make updates to protocols in real time. You may also want to consider moving...

...a programmable RNA-binding protein, creating the fluorescent fusion protein dLwaCas13a-NF for in vivo ...SpCas9 is not the only game in town, and other Cas proteins like SaCas9 and Cpf1 can circumvent the limitations...form of “bait” to identify new CRISPR-associated proteins within the bacterial genome. From this analysis...categories based on the architecture of the CRISPR protein in question: C2c1, C2c2 and C2c3 (short for Class...consistent with known roles for HEPN domains in other proteins. As with Cas9, mutating key residues in the Cas13a...is reduced when 2 mismatches are present. The protospacer flanking sequence (PFS) for LshCas13a, which ...cells. Indeed, groups have utilized dCas13 fusion proteins for imaging, tracking, modulating splicing, and...

...Fluorescent proteins (FP, e.g. GFP) can be used in many ways. Often a FP is fused to a protein of interest...leaf transformation: Check if there are working protocols available for your species This will save you ...first choice for establishing a new transformation protocol as it can easily show which tissue have been transformed...bioluminescence. Luciferases usually have shorter mRNA and protein half-lives then GUS and are therefore good temporal...elucidate the subcellular localization of your protein of interest. Find all plasmids from the Lee-Parsons...started. Once you have established a well working protocol in your lab, it can become a strong tool for your...plants using syringe agroinfiltration." Recombinant proteins from plants. Humana Press, 2009. 41-50. PubMed...

...Caused by a mutation in the methyl-CpG binding protein 2 (MECP2) gene, Rett Syndrome is a rare, progressive...Bird lab found that only a portion of the MECP2 protein is necessary to rescue the neurological defects...Goldilocks” principle, in which too much of the MECP2 protein is as detrimental as too little, resulting in MECP2...researchers is how to deliver the precise amount of protein to each brain cell to restore physiological levels...reactivation, RNA editing, RNA trans-splicing, and protein replacement to reverse the impacts of this disease...: Schematic of the human MECP2 locus and MECP2 protein, showing mutations corrected by CRISPR/Cas9 in ...in X-linked MECP2, encoding methyl-CpG-binding protein 2. Nat Genet 23:185–188 . https://doi.org/10.1038...

...length gene and/or protein is reconstituted. One way to reconstitute a split protein like Cas9 is to use...polypeptides that, when at the ends of two proteins, mediates protein trans-splicing, similar to an intron ...In search of a smaller, but equally potent Cas9 protein, the Zhang lab took a different approach. They ...toxicity. More AAV-based CRISPR systems Smaller Cas proteins SaCas9 isn't the only CRISPR enzyme that’s small...advantage that it can be turned off by anti-CRISPR proteins. The Sontheimer lab used NmeCas9 with AAV to edit...disease-causing loci in mouse liver.  Two other smaller Cas proteins include Cas12b and CasX. While there are no papers...papers yet published that deliver these Cas proteins with AAV, at just 1,108 and 986 amino acids, respectively...

...nuclease folds found in other proteins. The C-terminal domain (CTD) of the protein also belongs to the NUC ...important to choose a target sequence with a nearby protospacer-adjacent motif (PAM)? Cas9 is becoming an ever...lobe—do not structurally resemble any other known proteins; they are completely unique to the CRISPR system... a single unpaired strand in the center of the protein. The repeat:anti-repeat duplex pokes out from the...RNA, confirming its sequence. This region of the protein varies between Cas9 enzymes from different species.... The stem loops bind along the outside of the protein, spanning both lobes. Stem loop 1, nestled between...shows the guide RNA stabilizing the DNA-binding protein portion of the CTD, ready to check for PAM sequencines...

...sequence using all six open reading frames UniProt: A database of protein sequence and functional information...Biology Reference Find molecular biology protocols Browse our protocol video collection ... of Blue Flame plasmid pET21a-BirA.   DNA / protein database search and analysis tools Addgene's analyze...with Addgene. NCBI BLAST: Compares nucleotide or protein sequences to sequences in a database ExPASy translation...IHC.  FPbase: A data repository for fluorescent proteins and their properties.  Expasy: Bioinformatics ...needs 5 great apps for lab life  Sharing your lab protocols using apps to save you time and track your experiments...

...in hosts carrying the bacteriophage lambda pir protein. This allows conjugation between a pir-containing...might be useful in a broad array of pathogenic Proteobacteria. We placed the toxin genes under control of...that encode one of two visible blue or magenta chromoproteins (Figure 2) (Liljeruhm et al., 2018). This system...PMC3936740.  Giacalone, Matthew J., et al. "Toxic protein expression in Escherichia coli using a rhamnose-based...genome with two-step allelic exchange." Nature protocols 10.11 (2015): 1820. PubMed PMID: 26492139. PubMed...Exchange Vectors for the Scarless Modification of Proteobacterial Genomes." Applied and environmental microbiology...et al. "Engineering a palette of eukaryotic chromoproteins for bacterial synthetic biology." Journal of...

...dyes for DNA-protein interaction studies? Kyubong Jo's lab generated fluorescent protein fusions with ...DNA. TALE proteins can bind specific DNA sequences which brings the attached fluorescent protein to the ...contributed by Leah Schwiesow Calcineurin is a protein phosphatase that responds to rising intracellular... in the cell, calcineurin dephosphorylates the protein CZI, which in turn binds to calcineurin-dependent...10.15698%2Fmic2020.04.713 Truncated TALE-fluorescent protein fusions as a DNA staining dye Article contributed...stranded DNA bound by tTALE-FP with the fluorescent protein fluorescing green. Image from Shin et al., 2019...

...RNA. Argonaute proteins are also present in prokaryotes, and some of these proteins protect against invading...Biologists are going gaga over the newest gene-editing protein - a DNA-cleaving Argonaute from Natronobacterium...reasons why you might choose NgAgo over CRISPR proteins Cas9 or Cpf1 - keep reading to learn more! Background...: Argonaute and NgAgo In eukaryotes, Argonaute proteins play an important role in RNA interference, binding...Thermus thermophilus (TtAgo), showing that this protein could cleave DNA in vitro, directed by phosphorylated...loading appears limited to the period when the protein is being synthesized, so NgAgo remains faithful...

...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):...

... 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...

...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...

...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...