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Showing: 81 - 100 of 363 results
  1. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    ...protein. The prime editor consists of a Cas9 nickase fused to a reverse transcriptase. The Cas9 nickase is ...strand. The first iteration of prime editor simply fused the wild-type Moloney Murine Leukemia Virus (M-MLV...thermostability. What came out on top? The Cas9 nickase fused to a pentamutant of M-MLV RT. They called this system...3′ tail of pegRNAs (Yan et al., 2024). The team fused La to the C-terminal end of PEmax to produce PE7...pegRNA stability ~ ~ ~ ~ ~ ~ ✖ Fuse RNA-binding protein La to the prime editor Improve...
  2. 22 Hot Plasmid Technologies from 2014

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    Blog Post
    ...demonstrate that they can successfully create GFP fusion proteins with a variety of genes across the genome...dehydrogenation). MitoTimers are composed of a Timer protein fused to the mitochondrial targeting sequence of cytochrome...perform multiple site mutagenesis and create complex fusion or recombination vectors. The efficient and flexible... is a monomeric, 27 kD genetic tag which can be fused to proteins of interest for EM imaging, or targeted...colleagues, FokI-dCas9 expresses Fok1 nuclease domain fused to catalytically inactive Cas9 DNA-binding domain...single reaction, while retaining only a defined 4bp fusion site in between. The MoClo system is comprised...
  3. Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification

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    Blog Post
    ...domain.  How do these components interact? When fused together to form an OptoNB, the AsLOV2 domain acts...Since Ras is a membrane localized GTPase, the team fused a mCherry OptoNB to the catalytic domain of the ...that activates Ras. They expressed this OptoNB-SOS fusion protein in cells that expressed membrane-localized...activity Read about RANbodies, reporter nanobody fusions Learn more about using the secondary nanobody toolbox...
  4. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

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    Blog Post
    ...its target.  They name their most potent Acr-LOV fusion protein inhibitor CASANOVA for “CRISPR-Cas9 activity...works with the S. pyogenes Cas9, dCas9-effector fusions, and xCas9, and requires no additional chemicals...plasmid tools which includes plasmids encoding a fusion between cPYP and tgRFP (PLL7.0 tgRFP cPYP), and...or dark- state of the photoswitchable proteins) fused to mVenus linked to a mitochondrial localization...
  5. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

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    Blog Post
    ... Early efforts tested whether or not FP-tubulin fusion proteins could properly form microtubule networks...networks in cells. Other efforts tested whether FP fusions to connexin localized correctly to gap junctions...OSER assay with a number of fluorescent proteins. Fusions of CytERM with mTurquoise2 and dTomato, which are... localization and biological properties of new fusions can be compared with mEGFP or mTurquoise2, which...
  6. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

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    Blog Post
    ... cytoplasmic contents. Autophagosomes that have fused with the vacuole or a lysosome are trickier to identify... are degraded or released once an autophagosome fuses with the vacuole or lysosome. Rosella fluorescent...They are connected by a 9 amino acid linker that’s fused to the C-terminus of DsRed.T3. See Table 1 for a...Sargsyan et al. It is also possible to make Rosella fusions and thereby monitor autophagy of your protein of...
  7. What's New in CRISPR - November 2021

    Type
    Blog Post
    ... delivery into mammalian cells, especially when fused to effector proteins for gene activation or gene...the new system was able to activate expression by fusing dCas12f to VP64. Read the paper in Nature Biotechnology...combines an endogenous Gag homolog, cargo RNA, and fusogen in a modular system for delivering gene editing...active Cas9 enzyme. PE-Cas9 comprises an active Cas9 fused to the reverse transcriptase (RT), which induces...
  8. Viral Vectors 101: Voltage Indicators

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    Blog Post
    ... the second type of these GEVIs: a FP-fused rhodopsin. FP-fusion to rhodopsin resulted in electrochromic... Simply put, VSD-based sensors consist of a VSD fused to a single fluorescent protein (FP). This class...imaging.   Figure 1: Ace-mScarlet is a fusion between the voltage-sensitive inhibitory rhodopsin...
  9. Tips for Using BLAST to Verify Plasmids

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    Blog Post
    ...exact entry for your favorite gene fused to an epitope tag or fusion protein. If your sequencing primer...An alignment omitting repeated regions can be confusing, such as when you attempt to verify the starting...primer was chosen to confirm a tag or fusion protein is in-frame, we recommend using blastx with the "Align...
  10. Which Fluorescent Protein Should I Use?

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    Blog Post
    ...ectopically expressed in most organisms. Furthermore, fusing FPs to another protein usually does not affect ...and 2) to visualize the localization of the FP (fused to the protein of interest), thus tracking the localization...used to determine the proximity of two proteins fused to the halves of the split FP. This technique is...This may affect the biological function of the FP-fusion protein. Therefore, it is recommended to use monomeric...
  11. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...has focused on transcriptional activation using a fusion to a transcriptional activator such as VP64 (Gilbert...transcriptional activators (Qi et al., 2013). However, the fusion of dCas9 with a protein tag allows for the isolation... DNA fragment (Fujita et al., 2016). 3xFLAG-tag fused to dCas9, along with a biotinylated gRNA, were successfully... (Liu et al., 2017). This system utilizes dCas9 fused to a biotinylation tag expressed on a plasmid, allowing...
  12. Hot Plasmids - November 2023

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    Blog Post
    ...modular approach combines an interchangeable set of fusogenic and targeting components to reprogram delivery...Modular features of DIRECTED particles where the fusion and targeting components are interchangeable. Image...targetable cell types. DIRECTED is compatible with fusogens from different viral families and can be used ...transcriptional history of a cell. Lin et al. generated a fusion protein called iPAK4 that assembles into a slow-growing...
  13. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...MagIC-cryo-EM, target proteins (expressed as a GFP fusion in this proof-of-concept study) are captured on...programmable epigenetic editor consisting of dCas9 fused to a DNA methyltransferase — that can drive lasting...silencing, which can be toxic, CHARM uses a cofactor fused to a histone tail to recruit and stimulate the endogenous...multiplexed targets.   Figure 4: CHARM uses a fusion of the DNMT3L cofactor (D3L) with a histone tail...
  14. Antibodies 101: Flow Cytometry

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    Blog Post
    ...expression, measured using fluorescent protein fusions In addition to visible light, flow cytometers can...Which Fluorescent Protein Should I Use? However, fusion proteins tag your protein of interest with a fluorescent...- or C-terminus of the protein. One downside of fusion proteins is that the tag can alter the structure...get around the challenges of fluorescent protein fusions by binding to the protein in its native state.  ...
  15. Special Delivery: Fluorophore Targeting for FRET Studies

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    Blog Post
    ...experiments. However, remember that, if you make an FP fusion, you are inserting about 20-25 kDa of protein mass...to this bulky insertion are inevitable. Most FP fusions for FRET studies are at the N- or C-terminus of... protein of interest, resulting in a C-terminal fusion of a tetracysteine tag. In addition, since these...FRET Browse Empty Plasmids to Fluorescent Protein Fusions Check out Our Fluorescent Protein Guide ...
  16. Plasmids 101: How to Name Your Plasmid in 3 Easy Steps

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    Blog Post
    ... any tags or fusions that are on your insert. Typically you would list any tag or fusion protein in the... pBACKBONE-Flag-hGene If there was also an EGFP fused to the C-terminal of your insert you would list ...
  17. What's New in CRISPR - May 2019

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    Blog Post
    ...After testing dCas9 fusions with transcriptional repressors, they found that fusing dCas9 to an Mxi1 repressor...covalently attached to the Cas9-gRNA RNP complex via the fused HUH endonuclease PCV. This increases HDR efficiency...
  18. Inntags: Innovative Protein Epitope Tagging

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    Blog Post
    ..., Georgieva et al. examined the effects of GFP fusions to inntags or traditional FLAG, HA, and Myc tags...untagged GFP overexpression, while the rest of the fusions had some negative effect on these parameters. IT5... cell morphology was observed with IT5 and IT6 fusions. In contrast, Flag and HA tags had especially detrimental...
  19. Your Top Requested Plasmid in 2016!

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    Blog Post
    ...from the Zhang Lab. It's name, though a little confusing to the uninitiated, contains much of the information...directed by a gRNA. 2A-Puro In this plasmid, SpCas9 is fused to the puromycin resistance gene through a 2A peptide...the 2A peptide on its C-terminus, SpCas9 is also fused to a 3X FLAG tag at its N-terminus. This tag can...
Showing: 81 - 100 of 363 results