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We narrowed to 879 results for: IND

Showing: 41 - 80 of 879 results

  1. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    Published
    March 28, 2024, 1:15 p.m.
    ...attachment. The five other IPI rAbs bind RGD-binding integrins at non-ligand-binding sites and do not interfere...morphology changes induced by integrins binding the RGD motif compared to those binding the Leucine-Valine-Aspartate-Proline...integrin antibodies exist, almost all bind outside the ligand-binding pocket, their sheer size impeding ligand... investigators inhibited other RGD-binding integrins, individually and in combination, showing that among...study. Cells often express multiple integrins that bind the same ligand, making it difficult to dissect ...The collection includes 26 unique antibodies that bind discrete subunits or particular subunit pairs. Some...antibodies in the collection are able to block ligand binding — and therefore interrupt integrin function — providing...

  2. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    Published
    Dec. 6, 2022, 2:15 p.m.
    ...SF-iGluSnFR. Image from Aggarwal 2022.   Find iGluSnFR3 plasmids here! Find viral prep here: pAAV.hSyn.FLEX.iGluSnFR3...al. 2022, under CC-BY-NC license.   Find JEDI-2P plasmids here! Find viral preps here: pAAV-EF1a-DIO-JEDI...this in mind, Ed Boyden and colleagues set out to engineer a potassium ion biosensor with a binding affinity...of potassium binding proteins, they developed a family of fluorescent potassium indicators with a range...new tools to measure their favorite targets and finding new targets to measure! Let’s take a look at some...adapted from Ino et al. 2022, under CC-BY license.   Find MTRIA sensors here!   Ino D, et al. A fluorescent...doi: 10.1038/s41592-022-01597-x. New glutamate indicator iGluSnFR3 Improving on the glutamate sensor iGluSnFR...

  3. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    Published
    Dec. 20, 2017, 2:33 p.m.
    ...transferred to an industry position. If you cannot get an Industry H-1B, remember that industry career paths...Subject Industry H1B Academic H1B National Interest Waiver EB-1A EB-1B Cap subject industry H-1B There...process or quota like industry, these organizations can be a good choice for individuals needing sponsorship...importance of the individual’s work in their field and the impact of the individual’s past work on the ...passionate about working with scientific individuals to find exciting growth opportunities in life sciences...growing shift away from employers sponsoring individuals for green cards because of the cost, long timeline...available to you including careers in academia, industry, non-profit, and government sectors.  Within these...

  4. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ...canonical base editing window, base editing has higher efficiency and fewer indels than prime editing. But...increase activity, enhance binding between the template and primer binding site, increase processivity...nicking increases indel formation slightly. Designing the sgRNA with a spacer that only binds the edited strand... the indicated tool. ~ reflects an innovation that may optionally be combined with the indicated tool.... advances in this evolving field! Find prime editing plasmids! Find plasmids from David Liu's lab! This...developed base editors with a <8 base-pair editing window cannot edit within what Fyodor Urnov refers to ...edit all the target bases within the base editing window, while prime editors make a specific edit defined...

  5. Antibodies 101: Validation

    Type
    Blog Post
    Published
    March 24, 2022, 1:15 p.m.
    ...too common for antibodies to bind to off-target molecules or even to not bind to their target molecule at... to make sure that your antibody binds to what you expect it to bind to. Show specificity for the target...because an antibody can bind to its target, does not mean that is all it will bind to - particularly when...of antibody-independent approaches to your antibody-dependent approach. Antibody-independent methods may...different tissues or cell types. If your antibody indeed binds to your target then you would expect to see ...influence antibody binding by altering the relative concentration of potential antibody binders and so this ...of the researcher toolkit due to their ability to bind to molecular targets with high affinity and specificity...

  6. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    Published
    May 8, 2018, 1 p.m.
    ... cut site.     Indel Distribution In the Indel Distribution tab, you’ll find an Indel plot (left panel... 21+ bp indel. This score is a useful measure to determine how many of the contributing indels are likely...deletions (indels) and then characterizes the sequence and abundance of each particular indel.   For a ...processing, the window shows a yellow checked circle. Typically, a yellow check mark indicates that a particular...will open up a new window with three tabs.  Each of the three tabs - Contributions, Indel Distribution and...distribution of indel sizes in the entire edited population of genomes. Hovering over each bar of the indel plot ... Check out our CRISPR guide Find CRISPR plasmids for your research Find other CRISPR software resources...

  7. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

    Type
    Blog Post
    Published
    April 23, 2018, 2 p.m.
    ...make it easier for you to find and use the plasmids you need. Below you'll find our hot plasmid articles...chemically inducible approaches to provide temporal control, this new system, Fkbp/Frb Inducible Recruitment... GAL4 or ZIF268 DNA binding domains fused to either the hER or hPR ligand binding domains and the MSN2...expression is dependent only upon its cognate inducer. Since induction via estradiol and progesterone do not ...transcription factors in series and independently changes inducer levels. That is, if transcription factor...created stable cell lines overexpressing TRIM21, finding that TRIM21 overexpression did not affect cell ...changes in expression of TRIM21-associated genes, indicating that TRIM21 overexpression does not aberrantly...

  8. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    Published
    June 20, 2024, 1:15 p.m.
    ...protein DnaA. First, DnaA must bind ATP. Then, the ATP-bound DnaA binds multiple specific sites within...controlled plasmids. Binding of IHF (green) redistributes DnaA (red) from binding primarily high-affinity...protein binding it (Grimwade et al., 2018). Rather than serving as an energy source, ATP binding is one...interacts with DnaA, stabilizing its binding and contributing to DNA unwinding. This promotes initiation at the... plasmids that can initiate DNA replication independently are said to be under relaxed control, as they...come in both high- and low-affinity versions, but binding of the low-affinity versions is required to start...DnaA alone, as long as there are enough copies to bind every available DnaA box. In bacteria, however, ...

  9. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    Published
    Feb. 13, 2025, 2:15 p.m.
    ...arrows indicate a conversion directly catalyzed by the base editor. Dotted red arrows indicate the change...well as a 2.3 fold decrease in indel formation. To further decrease indel formation 1.5-2 fold, the team...editing window of target positions 4-7. ABEs 6.3, 7.8, and 7.9 display slightly wider editing windows of position...but higher indel formation than BE2 Nishida et al., 2016 Target-AID   Edits 3-5 base window surrounding...following base editing and DNA repair. Black arrows indicate natural cellular processes. BER = base excision...locus A target base for editing within the editing window specified by the Cas protein These elements were...of targets in human cell lines, with an average indel frequency of only 1.1%. These numbers are a vast...

  10. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ... make it easier for you to find and use the plasmids you need. You can find all the hot plasmids from ...SpCas9 (NGG)  Editing window ~2 nt  85175  pBK-EE-BE3  SpCas9 (NGG)  Editing window ~2 nt  85176  pBK-YE2...essential in B. subtilis 168. Each individual strain includes a xylose-inducible dCas9 paired with a single, ...found that TZAP binds directly to TTAGGG repeats via its zinc finger domains. This binding was also independent...independent of another TTAGGG binding complex, shelterin. Swapping the DNA binding domain of another telomeric...N-terminal variants on iPSC generation Deriving iPSCs (induced pluripotent stem cells) through the expression ...valuable tool for the research community. You can find the AAV vectors for infecting GABA-ergic interneurons...

  11. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    Published
    Sept. 21, 2017, 2:07 p.m.
    ...propensity of Cas13a to bind and cleave RNA non-specifically. The ability of dCas13a to bind target RNA without...function, and will therefore be the focus of the remainder of this blog post. Perhaps the biggest difference...difference between Cas13a and Cas9 is that Cas13a binds and cleaves RNA rather than DNA substrates. In terms...nuclease dead” Cas13a (dCas13a), that is capable of binding target RNA but lacks the ability to cleave the ...Cas9 uses both a tracrRNA and crRNA to facilitate binding and cleavage of target DNA respectively. LshCas13a...a uracil-rich stem loop and facilitates target binding and cleavage through a series of conformational...like Cas9 or Cpf1. As a result, Cas13a will then bind and cleave additional RNAs regardless of homology...

  12. A Control for All Seasons

    Type
    Blog Post
    Published
    Oct. 5, 2023, 1:15 p.m.
    ...antibody not only binds specifically to its target through the antigen binding site but also binds non-specifically...Preimmune serum control will not bind to the desired target but will bind nonspecifically through the Fc...detectable, it is likely due to binding interactions outside of the antigen-binding site.      Secondary antibody...blot and look for the molecular weight shifts indicative of tau phosphorylation. By including the positive... are likely just natural differences between individuals. Recommended controls for antibody-based experiments...secondary antibody cross-reactivity. Non-specific binding  Primary antibody specificity is further supported...

  13. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    Published
    Nov. 3, 2015, 3:30 p.m.
    ...computational support. Indeed, rational sgRNA design is not possible without relying on some kind of pre-analysis... needs of each individual; the  community as a whole has yet to settle on any true kind of value. I’ve... because they rely on pre built indices or databases in order to find and present the results to you as...this, some online tools such as Cas-OFFinder will suffice. Cas-OFFinder provides a bare-bones data dump of...offline front, there is a Python script called SSFinder, but I find its implementation to be too slow for practical... are altogether different. They are filled with blind anticipation. Their trust in the quality of the ...reviewing the contents of the piñata and the audience blindly receiving them; it results in highly similar content...

  14. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    Published
    Jan. 5, 2021, 2:15 p.m.
    ...sign up here.  Here's what you'll find in this post: Blue-light inducible Cre for use in bacteria CRISPR-Casɸ...ABE.  Find the plasmid here Jesse Zalatan’s lab deposited plasmids for a chemically inducible CRISPRa...deposits and we hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like...items from the viral service A fast blue-light inducible Cre for precise control of bacterial gene expression...of optogenetics. Optogenetic tools use light to induce conformational changes in a protein.    ...Tagging with Light (SPOTlight) as a way to isolate individual cells with unique spatiotemporal phenotypes from...a few highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene...

  15. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    Published
    Jan. 8, 2019, 1:54 p.m.
    ...this plasmid which indicates that there was a prior version of this plasmid. Indeed the first version ...second most requested plasmid of all time, pMD2.G. Find psPAX2. pMD2.G - Also from the Didier Trono lab ...This plasmid was the top requested plasmid of 2017. Find pMD2.G. pSpCas9(BB)-2A-GFP (PX458) - This plasmid...2A-GFP allows for screening of transfected cells. Find pSpCas9(BB)-2A-GFP (PX458). lentiCRISPR v2 - This...for CRISPR pooled library screening experiments. Find lentiCRISPR v2.(If you’ve peeked through the rest...oligos can be cloned to create a chimeric guide RNA. Find pX330-U6-Chimeric_BB-CBh-hSpCas9. pSpCas9(BB)-2A-Puro...the most recently available plasmid in the top 15. Find pSpCas9(BB)-2A-Puro (PX459) V2.0. pCMV-VSV-G - ...

  16. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...deposits and we hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like...deposit please sign up here.  Here's what you'll find in this post: Yeast-based protein evolution Fluorescent... proteins and might be useful for your studies.  Find the plasmids to set up your screening systems at...tagged with MapTau to label entire microtubules. Find these fluorescent mammalian cell structure markers... T7 Express competent E.coli (NEB) using IPTG induction. To biotinylate the nanobodies, the authors used... and results in uniform NB biotinylation. Once induced, the NBs could be easily purified by one-step immobilized...variations in human pluripotent stem cells(hPSCs), Lindy Barrett’s lab at the Broad Institute developed fluorescently...

  17. Plasmids 101: Control Plasmids

    Type
    Blog Post
    Published
    April 9, 2015, 3:29 p.m.
    ... correct interpretation of the effect of the independent variable you are testing. Importantly, they demonstrate...experiment is specifically associated with the independent variable you are testing and not some other unintended... are used to minimize any effects of the non-independent variables.  To illustrate the importance of control...control; specifically, the plasmid without the independent variable. Referring back to the experiment associated...associated with Figure 1, the independent variable is the shRNA. Therefore the empty control vector would... Plasmid B (as internal control) This result indicates that the transfection was not successful due to...including how much plasmid DNA to use for any individual or co- transfection, should be determined empirically...

  18. Deep Dive: Statistical Tests (Comparisons)

    Type
    Blog Post
    Published
    March 7, 2023, 2:15 p.m.
    ... that the independent variables are independent of each other. That is, multiple independent variables...the production window. Factorial ANOVA This type of ANOVA assesses multiple independent variables on a...distribution, equal variance, and independent variables), it’s best to find a test that does. If you perform...relevant for many downstream statistical tests.   Independent variables – factors which are not affected by...assumptions: Distribution – Normal Variables – Independent Variance – Approximately equal Data type - Quantitative...t-tests can also be referred to as two-sample or independent, but these designations all refer to the same... for example) simultaneously, with up to two independent variables. This test will tell you if there are...

  19. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    Published
    June 27, 2024, 1:15 p.m.
    ...design. Find streptavidin plasmids and biotin plasmids at Addgene!     Figure 1: Indirect detection...for example, a previously induced gene knockout. Besides amine- and DNA-binding dyes, nucleoside analogs...live cells are protected. Amine-binding viability dyes can still bind free amines on the cell surface,...where fluorophore-conjugated antibodies directly bind to epitopes of interest that are found in the extracellular...and 'staining' to refer to small molecule dyes. Indirect labeling While antibodies used for target marker...western blotting or ELISAs.  A popular method for indirect labeling employs a primary epitope-targeting antibody...fluorophore-conjugated streptavidin protein, which binds the primary antibody through the biotin-streptavidin...

  20. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    Published
    Nov. 16, 2021, 2:15 p.m.
    ... In contrast, indirect IF requires two antibodies: a primary and secondary. For indirect IF, the primary...immunofluorescence and indirect immunofluorescence work. Image created with BioRender.com   Indirect IF is a much...are applied to a cell or tissue sample where they bind to their targets, most commonly proteins. You can...molecules, knowing that where you see fluorescence indicates your target’s location. Some other terms you might...learn more about these techniques!  Direct vs. indirect immunofluorescence There are a couple different...familiar with when talking about IF, direct and indirect. For direct IF, researchers use a single antibody...on this technique here. One of the reasons that indirect IF is more common is that secondary antibodies...

  21. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    Published
    Feb. 25, 2016, 3:30 p.m.
    ...executive director of Central Indiana Science Outreach and a Postdoc at Indiana University School of Medicine...instance, I tweeted to the Indiana State Museum and others when I was trying to find local informal science... mine was geared toward careers in academia or industry. Luckily, I discovered that there are many ways...culminated in my founding a nonprofit called Central Indiana Science Outreach, or CINSO. We organize fun science... you’re provided throughout your PhD program.   Find people who will support your professional development...advisors are not as supportive of your career goals, find people who are. I wanted to do fun science activities...

  22. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    Published
    Oct. 19, 2017, 1:54 p.m.
    ...Resources on Addgene.org Find Cre-Lox Plasmids Find Optogenetics Plasmids Find Chemogenetics Plasmids ...recombinase together with inducible Cre recombinase (CreERT2), visualizes individual neurons and their synapses...but share an overall structure of 13 base pair palindromic repeats separated by an 8 bp asymmetric core.... recombinase efficiency are important to keep in mind when deciding whether to use the Cre-lox or one ...compatible target sites are cleaved out, leaving behind a vector with the inverted DNA flanked by non-compatible...networks Double inverted vectors expressing light inducible ion channels (plasmid # 20298 for example) have...to fuse a mutant estrogen receptor (ER) ligand binding domain (LDB) to the C-terminus of FLP or Cre (pCAG-CreERT2...

  23. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ... does inducible expression work? As mentioned above, many expression plasmids utilize inducible promoters...'inactive' until an inducer such as IPTG is added to the growth medium. Induction timing is important,...long to induce, your culture will start collecting dead cells, and, conversely, you cannot induce too early...compatible. An arabinose-inducible plasmid will not express in an IPTG induction strain for example, nor...engineered to accomodate high protein loads using inducible promoter systems (which will be discussed later...Features Genotype Use BL21 (DE3)   Basic IPTG-inducible strain containing T7 RNAP (DE3) F- ompT lon hsdSB...toxic proteins BL21-A1 Tetracycline Arabinose-inducible expression of T7 RNAP; IPTG may still be required...

  24. RaPID Detection of RNA-protein Interactions

    Type
    Blog Post
    Published
    Aug. 16, 2018, 12:40 p.m.
    ... lower IREB2 binding enrichment compared to wild-type sequences and that loss of binding correlated with...with Split-BioID Find resources for studying chromatin Resources on Addgene.org Find Split-BioID plasmids...plasmids Find CRISPR plasmids for labeling genomic regions Find CRISPR plasmids for purifying specific genomic...characterization of RNA-protein interactions has lagged behind, likely due to limitations of current means to ...and allows a researcher to identify proteins that bind an RNA motif of interest in living cells. Challenges... has limited utility for identifying novel RNA-binding proteins that interact with an RNA of interest....distinguish between proteins that directly or indirectly interact with the bait. Also, RaPID can’t be ...

  25. Antibodies 101: Fc Effector Functions

    Type
    Blog Post
    Published
    May 18, 2023, 1:15 p.m.
    ...antibodies to bind. We also know that changing the Fc domain can impact an antibody's binding affinity (Janda...express Fc receptors that bind to the Fc domain of an antigen-bound antibody and induce (or, in the case of ...interact with, so IgG isotypes bind to Fcγ receptors; IgE isotypes bind to Fcε receptors; and so on and...antibodies bind to a pathogen and natural killer (NK) cells expressing Fcγ receptors binds to IgG and ...know much about yet. Who knows what kinds of functions they may induce?  Fc receptors are intriguing from...system, including the innate immune response, by binding to Fc receptors on other immune cells. (psst! Need...Antibody-dependent complement deposition (ADCD) Antibodies can bind to a pathogen and physically prevent it from entering...

  26. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...(470nm), paCas9-1 induced indel mutations via NHEJ (frequency of 20.5%) and induced modifications by HDR...the light-inducible heterodimerizing cryptochrome 2 (CRY2) and calcium and integrin-binding protein 1 ...background and high fold-induction of Cas9 activity (16.4-fold). This paCas9-1 light-inducible system was able ...flanking the DNA-binding cleft of Cas9, the fluorescent domains dimerize and prevent DNA binding in the dark...Upon expression in the cell, the dCas9-CIB1 fusion binds to the target DNA sequence as directed by the guide...background activity in the dark state and highest fold induction at 31X. By using a slit pattern during blue light...domain VP64. Using this system in HEK293T cells to induce expression of human IL1RN, the researchers saw ...

  27. Design Tips for Prime Editing

    Type
    Blog Post
    Published
    Jan. 23, 2025, 2:15 p.m.
    ...primer binding site, starting with a length of about 13 nt (Anzalone et al., 2019). Primer binding sites...designed to bind only after the edit is installed. This reduces concurrent nicks, which lowers indel rates ...the primer binding site (Nelson et al., 2022). PE7: This editor includes the fused RNA-binding protein La... of a CRISPR gRNA extended to include a primer binding site (PBS) complementary to the genome and a reverse...Starting lengths for optimizing your pegRNA primer binding site (PBS) and reverse transcriptase template (...), disrupting its canonical structure and Cas9 binding (Nishimasu et al., 2014). Figure 3: pegRNAs...while pegRNAs that use other bases in this location bind to Cas9 with canonical sgRNA structure (right). ...

  28. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    Published
    Feb. 25, 2020, 2:15 p.m.
    ...reinforced by the findings of the 1952 Hershey-Chase experiment. In the same year, Rosalind Franklin and her...The DNA-binding domain can be engineered to target specific DNA sequences while FokI then induces a double...resolved the DNA double helix. In the 1960s, Har Gobind Khorana, Marshall W. Nirenberg, and Robert W. Holley...2018) were highly successful in the box office, indicating the insatiable appetite of the moviegoers for... enzymes generated by fusing a zinc finger DNA-binding domain to a DNA-cleavage domain (FokI). Zinc finger... (TALENs) were engineered. TALENs follow a DNA-binding and DNA-cleaving strategy similar to the ZFNs, ...nucleotide triplets, the components recognize individual nucleotides. This imparts TALENs more specificity...

  29. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ...oligos that bind with high affinity to small molecules to induce their fluorescence. Until a binding event ...nonspecific binding with RNA by about 7x). Mango fluoresces nearly as brightly as Spinach, but binds with much...small molecule are strongly fluorescent but the binding of an aptamer to its target small molecule activates... process used to evaluate oligonucleotides for binding affinity to a specific target, and the molecular...Broccoli and their derivatives are aptamers able to bind to a small molecule derivative of GFP called 3,5...Additionally, Spinach, the first aptamer identified to bind DFHBI, is highly unstable in vivo due to poor folding...naturally occurring regions in the 5’ UTR of mRNA that bind metabolites, leading to conformational changes that...

  30. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ... - all without the need to find a llama farm! To find an optimally binding sybody, you want to start out...COVID-19. Researchers identified nanobodies that bind to coronavirus spike proteins (Wrapp et al., 2020...days and isolated special camelid antibodies that bind to the spike proteins in cell culture, and inhibit...small (12-15 kDa) single-domain proteins that can bind effectively to protein targets, similar to how antibodies... “arms” holding the highly variable regions that bind to specific antigens. In contrast, nanobodies are...of sybodies that hypothetically contain ones that bind strongly to your target protein. The Seeger lab ...corresponding to different amino acids) in the regions that bind to the antigen. The Seeger lab can directly provide...

  31. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...stringency of your binding. And most importantly: Independently validate identified binding partners with ...the DNA binding domain (DBD) and transcriptional activation (TA) domains retain their individual activities... as using five independent protein interaction detection methods (6). In order to bind DNA and activate...successfully expressed as fusions that retain their individual activities (1). Researchers also discovered the...DBD of the fusion protein shown is still able to bind to the upstream activation sequence (UAS) of its...gene then allows detection of those cells where binding between the proteins of interest has occurred. ...histidine-auxotrophic host strains are used to select for binding-dependent expression of the HIS3 gene, which is...

  32. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    Published
    June 24, 2020, 5:45 p.m.
    ...modifiers to specific loci without inducing double strand breaks. Below you'll find some of the CRISPR-based epigenetic..., KRAB induced silencing was transient and quickly reversed in culture. However, DNMT3A-induced methylation....2016.08.056 Find plasmids from this publication at Addgene. Maeder ML, Angstman JF, Richardson ME, Linder SJ,...catalytic domain, could mediate demethylation and induce transcription at CpG regions of various promoters...modifiers, and is available as a kit from Addgene.   Find Ready-to-Use Viral Preps for Epigenetic Modification...experience with these constructs in the comments! Find Plasmids for Epigenetic Modification   Leah Schwiesow...Biotechnology 33:510–517 . https://doi.org/10.1038/nbt.3199 Find plasmids from this publication at Addgene. Klann...

  33. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    Published
    Dec. 7, 2021, 2:15 p.m.
    ...beads and allowed to bind to the antibody being used for IP. These proteins bind only to the Fc portion...Protein G will bind both the primary antibody and serum immunoglobulins indiscriminately, causing competition...triangles. 2) An antibody immobilized on a bead binds specifically to the blue triangle protein but not... During the incubation the immobilized antibody binds to the target protein, thereby tethering it to the...method is desirable if the antibody is expected to bind weakly to the target protein or if the target is... beads in a way that also allows the antibody to bind to the target protein. One of the most common immobilization...cell wall proteins, Protein A and Protein G, which bind to the fragment crystallizable region (Fc) of antibodies...

  34. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    Published
    April 3, 2014, 8:05 p.m.
    ...transcribed promoters Inducible with galactose; repressible with glucose  Can be used independently or together....not induced. Requires T7 RNA polymerase, which is also controlled by lac operator. Can be induced by IPTG...presence of glucose or by competitive binding of the anti-inducer fucose Weaker. Commonly found in pBAD...the combination of the promoter (RNA polymerase binding site) and operators (response elements). Promoters...The sequence of the promoter region controls the binding of the RNA polymerase and transcription factors...specificity, the eukaryotic RNAP can recognize and bind to specific promoter elements. This means that the...expression mRNA Tetracycline response element promoter Inducible with Tetracyline or its derivatives.  Typically...

  35. What's New in CRISPR - September 2019

    Type
    Blog Post
    Published
    Sept. 10, 2019, 12:56 p.m.
    ...mechanism behind its activity including an extensive RNA scaffold and a domain needed for DNA unwinding. They...expression. Read the paper in Nature Find the CasX plasmids Drug inducible transcriptional activation and ... the paper in Nucleic Acids Research Find the drug inducible Cas9 plasmids E. coli genome-wide inhibition...suppressor genes (TSG). Find the Mouse Metastasis CRISPR-Cpf1 Double Knockout Library Find the Mouse Tumor ...Addgene.org Read our CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ... but we hope that this blog series will help you find new CRISPR tools for your research! This time: ...GeneWeld vectors to create knock-ins CasX Drug inducible CRISPR/Cas9 activation E. coli genome-wide inhibition...

  36. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    Published
    May 18, 2021, 1:15 p.m.
    ...it easier for you to find and use the plasmids you need. Here's what you'll find in this post: Pooled... possible amino acid mutations of the receptor binding domain (RBD) of the viral spike protein.   ...2021. SARS-CoV-2 infects host cells through binding the human angiotensin converting enzyme 2 (ACE2...receptor. The spike protein RBD responsible for binding ACE2 is the target of most anti-SARS-CoV-2 antibodies...RBD mutations which do not interfere with ACE2 binding, but impact recognition by specific anti-SARS-CoV...characterized, and validated by the Gräslund lab. Recent findings suggest that aaRSs are involved in inflammation...fluorescence, and suspension bead assay (Figure 2). After finding no cross-reactivity, antibody candidates underwent...

  37. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    Published
    May 30, 2019, 12:44 p.m.
    ...Addgene.org Read our CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...dCas9 (Jinek et al., 2012). Like Cas9, dCas9 can bind to a specific DNA sequence via a targeting gRNA.... 2013), or repression of transcription through binding a promoter region to inhibit association of transcriptional...Fujii, 2013). They call this method engineered DNA-binding molecule-mediated chromatin immunoprecipitation...immunoprecipitation (enChIP) because any engineered DNA-binding molecule, such as Zinc-finger or TAL proteins, can be used...genomic DNA that could then be recognized by a DNA-binding molecule (Hoshino and Fujii, 2009). Other forms...study. In addition to the identification of DNA binding proteins, this method can characterize nucleic ...

  38. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...the mitochondria (binding FRB), allowing for a rapid and inducible inactivation. Find the Robinson plasmids...that can rapidly induction heterodimerization of proteins that contain rapamycin-binding domains, such as...mitochondrial matrix. Gottlieb’s lab has used an inducible MitoTimer reporter using a Tet-on system (pTRE-Tight-MitoTimer...distinct differences of red-to-green fluorescence in individual mitochondria in the networks marks the differences...protein may upregulate an alternative pathway or induce other compensatory mechanisms. How can protein ... as FKBP and FRB. The cell does not appear to mind extraneous proteins bound to its outer mitochondrial...protein with a FRB domain (MitoTRAP). Rapamycin binds your recombinant protein (via FKBP) and then sequesters...

  39. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    Published
    May 23, 2023, 1:15 p.m.
    ...antibody binds a target protein in your sample, then a fluorescently labeled secondary antibody binds that...like. Plus, indirect IF amplifies the signal since multiple secondary antibodies will bind to each primary...Figure 1A), you’ll block, add primary antibody to bind the target, wash, add dye-conjugated secondary antibody...there are a few specific considerations to keep in mind. Most importantly, make sure you are using primary...However, the primary-secondary workflow (a.k.a. indirect IF) is more flexible and efficient, since you ...antibodies that have been validated in IF applications (find IHC validated or ICC validated antibodies from Addgene...and validate the antibody yourself. If you can’t find a suitable antibody against your target protein,...

  40. What's New in CRISPR - December 2019

    Type
    Blog Post
    Published
    Dec. 17, 2019, 2:37 p.m.
    ...in Nature Biotechnology Find the base editors   Single base editor that induces C-to-T and A-to-G mutations...sgRNA scaffold with a com aptamer which binds an aptamer binding protein for packaging into a lentivirus-like...Addgene.org Read our CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ... but we hope that this blog series will help you find more new CRISPR tools for your research! This time...Expression of this system is tunable via a rhamnose-inducible promoter. While this system was optimized for ...used in. Read the paper in ACS Synthetic Biology Find the CRISPRi toolkit SECURE adenine base editor...editors into one protein. They fused an activation-induced cytidine deaminase (AID) and an engineered tRNA...
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