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We narrowed to 612 results for: SCR

Showing: 101 - 150 of 612 results

  1. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    Published
    Dec. 6, 2022, 2:15 p.m.
    ... They use a unique and powerful high-throughput screen to select variants based on performance metrics...: JEDI-2P was identified from a high-throughput screen, selected for increased voltage response, brightness...a new preprint, Jared Toettcher and colleagues describe pYtags, a modular system for monitoring specific...developed using a high-throughput multiparameter screen (Koveal et al. 2022). Lactate is a key metabolite...cellular concentrations of lactate. The microfluidic screening platform developed here is also a promising approach...Figure 6: A) Schematic of BeadScan platform for screening candidate biosensor variants for performance under...Koveal D, et al. A high-throughput multiparameter screen for accelerated development and optimization of...

  2. Cellosaurus: A Cell Line Information Database

    Type
    Blog Post
    Published
    Feb. 21, 2023, 2:15 p.m.
    ...database describing nearly 145,000 cell lines. Cellosaurus Cellosaurus seeks to identify and describe all ... resource.  This is a labor-intensive project; scripts can be used in some cases, but much of this work...authors of the paper in which the cell line was described. Using Cellosaurus Cellosaurus is searchable by...information about a cell line you’ve used? Submit your manuscript or data to Cellosaurus via [email protected] received a report that HT-55, which was described as a colon cancer cell line, is actually a rectal...

  3. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

    Type
    Blog Post
    Published
    July 7, 2015, 3:36 p.m.
    ...have used PCR-based screening to detect mutants, but this method limits screening to a few hundred worms... the summer of 2013, a remarkable nine papers describing CRISPR/Cas9 genome engineering methods for C....can generate a fluorescent protein fusion, transcriptional reporter, and loss-of-function allele in just...knock-ins. These candidate animals can then be further screened based on their Roller phenotype. If SEC is inserted...wild-type animals that have undergone SEC excision are screened based on the loss of the Roller phenotype. These...

  4. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    Published
    March 6, 2018, 1:49 p.m.
    ... have been generally found to have a higher discrimination at the upstream side of the ligation junction...including Taq DNA ligase, are naturally able to discriminate against ligating substrates containing base ... at elevated temperatures, allowing further discrimination by incubating the ligation at a temperature... 27). This methodology has been used to rapidly screen buffer conditions for Taq DNA Ligase and their ...electrophoresis (CE) (see below). This method allowed screening of all possible base combinations (Watson-Crick...of a 96-well plate, allowing 6 conditions to be screened per plate. The results indicated that the optimal...optimization method has been used internally at NEB to screen additional ligases, conditions, and formulations...

  5. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    Published
    March 7, 2017, 3:30 p.m.
    ...Zhang and is described in Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE,...is bad news for repeatability and bad news for a screen. Because Addgene viruses are provided at a specific...expression cell line. Especially if you’re doing a screen. I made a few monoclonal lines (Figure 2) and I...was a gift from John Doench & David Root and is described in Rational design of highly active sgRNAs for...were gifts from John Doench & David Root and are described in Optimized sgRNA design to maximize activity...Additional Resources on the Addgene Blog Genome-wide Screening Using CRISPR/Cas9 Deliver Multiple gRNAs at Once...

  6. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    Published
    Oct. 26, 2017, 1:59 p.m.
    ...analysis scripts available, some associated with specific libraries, but any analysis script should return...plasmid. However, lots of library DNA is needed for screening experiments, so the scale of the DNA prep is larger... mixture on solid media. The colonies are then scraped off the plates and pelleted for DNA extraction....automated manner. Addgene commonly uses a custom script derived from the count_spacers.py file from Feng...wishes you luck in your future library use and screening, and if you have any tips, input, or stories of...

  7. Addgene Welcomes John Doench to the Advisory Board

    Type
    Blog Post
    Published
    Aug. 13, 2015, 2:30 p.m.
    ... primary screen to knowing what are the real hits is so much shorter…. it’s a great screening technology...researchers in a variety of fields develop RNAi screens to answer their own research questions. Starting... For his post-doc, Doench worked on using RNAi screens to learn more about cancer biology in Ed Harlow...developing CRISPR pooled libraries for use in genetic screens and has contributed much of his knowledge to the...become more efficient in terms of editing and screening) such that thousands of individual edits can be...

  8. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    Published
    June 2, 2015, 6:06 p.m.
    ...tRNAs contain internal transcriptional elements that enhance pol III transcription, potentially increasing...base Csy4 recognition sites from a polycistronic transcript. Although this method shortens construct size...coexpression of Csy4 with Cas9 and the polycistronic transcript. As nucleases may have off-target effects and...matching traditional gRNA-expressing constructs, transcript levels of PTG1 and PTG2 were 3 and 31 times higher...worries, I've summarized the three strategies described in this post in the graphic below - including ...

  9. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ...system is much more potent than the in vitro-transcribed single guide RNA (IVT sgRNA) approach (Dewari...comparison of Cas9 RNPs prepared with an in vitro-transcribed (IVT) sgRNA versus synthetic cr:tracrRNA. Cells...epitope-tagging experiments. Notably, we could tag >60 transcription factors in a short (3-week) time window; this...epitope tagging experiments: Retrieve Ensembl Transcript ID(s) of your favourite gene(s) from Ensembl .../index.html Design crRNA/ssODN: Using Ensembl Transcript ID(s) as an input query, pick crRNA (guide RNA...strand (strand used by RNA polymerase II for transcription) are more effective for genome-editing events...step by step protocol for the csRNP method is described in our recent article (Dewari et al. 2018), however...

  10. RaPID Detection of RNA-protein Interactions

    Type
    Blog Post
    Published
    Aug. 16, 2018, 12:40 p.m.
    ...and IREB2, which post-transcriptionally regulate iron metabolism pathway transcripts. By using RaPID, Ramanathan...signal (~250-fold enrichment) when compared to a scrambled RNA motif control). BioID2 had no detectable signal...experiments RaPID negative controls include both scrambled RNA motifs and cells transfected with BirA* protein...changes in RC2H1 protein expression and post-transcriptional regulation of SM1v1-containing RNAs could play...IRE) RNA motif of the ferritin light chain RNA transcript. IRE motifs are bound by IRE binding proteins...

  11. dTAG - You're it!

    Type
    Blog Post
    Published
    June 21, 2018, 2:06 p.m.
    ...locus-specific knock-in of FKBP12F36V. Detailed protocols describing the use of these plasmids are available here....example, IZKF1 and IZKF3 are cell-type specific transcription factors with important roles in Multiple Myeloma... (Nabet et al., Erb et al., Huang et al.), transcriptional and epigenomic profiling studies (Nabet et ...degradation, and lead author on the recent paper describing the dTAG technology. Follow him on twitter @behnamnabet... 29581585. This reference contains the full description of the dTAG system. The dTAG system was employed...Featured molecules: dTAG-7 and dTAG-13 Erb et al. Transcription control by the ENL YEATS domain in acute leukemia...

  12. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    Blog Post
    Published
    March 12, 2020, 1:15 p.m.
    ...synthetic promoters. Legend: TSS: transcription start site. TFBS: transcription factor binding site. Image from..., the Roska lab built synthetic promoters from scratch and tested to see if they drove cell type specific...conserved sequences that are upstream of the transcription initiation sites of at least two genes with ...promoters by repeating mouse cell type specific transcription factor binding sites interspaced with random...designed using stretches of DNA that were transcriptionally active in retinal cell types in mice. ...

  13. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    Published
    Dec. 14, 2021, 2:15 p.m.
    ...affect transcription independent of their enzymatic activity. They interact with transcriptional machinery...specifically edit methylation sites.  Step aside, transcriptionally active dioxygenase  When I started to work...by distinct proteins and result in unclear transcriptional consequences (Sprujit 2013). Enzymatic domains...in the genome and directly interacting with transcription machinery.   Well, I’ve got some good news...Hydroxylation, Active DNA Demethylation, Gene Transcription, and Memory Formation. Neuron 79, 6 P1086-1093...

  14. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    Published
    March 8, 2017, 3:15 p.m.
    ...spreadsheet Data Field Description Plasmid Name Use a descriptive plasmid name and match the... entry process On the Addgene home page, if you scroll over “Deposit” in the mega menu at the top of the... in red in the spreadsheet (Figure 2) and are described in Table 1. Some cells have drop-down options ...does this plasmid do?” in <200 characters. A descriptive purpose focuses on the practical application ... Primary Vector Type The vector type should describe how your plasmid is intended to be used. Choose...

  15. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ...their coat protein to screen for binding to the target protein. The protocol describes going through two ...protein (sybody), ribosome, and mRNA. During the screening process, displayed sybodies that bind to the biotinylated...enriched pool of sybody candidates are then reverse transcribed into cDNA fragments that are efficiently subcloned...The enriched sybody pools from the phage display screens are subcloned (using FX cloning) into the expression...interest. The Seeger lab used the ELISA technique described in their protocol because it is less susceptible...

  16. Common Injection Routes in Mice

    Type
    Blog Post
    Published
    Jan. 22, 2025, 9:42 p.m.
    ...off-target pokes to nearby organs. Restraint: scruffing Location: peritoneal cavity Gauge: 25–27 Volume...suspension to create xenograft models. Restraint: back scruffing or mechanical restraint Location: back or hind...but has a pretty low learning curve! Restraint: scruffing Location: throat Gauge (bulb-tipped feeding tube...to intermediate (tail vein) Intraperitoneal Scruffing Peritoneal cavity 25–27 < 2–3 mL Intermediate ... < 0.02–0.05 mL Beginner Subcutaneous Back scruffing or mechanical restrainer Back or hind legs; between...Anesthesia Nose n/a < 0.05 mL Beginner Oral gavage Scruffing Throat 18–20 (bulb tipped) up to 10 mL/kg Intermediate...

  17. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    Published
    June 25, 2024, 1:15 p.m.
    ...typically a Cas9 H840A nickase fused to a reverse transcriptase) together with a prime editing guide RNA (pegRNA...containing a primer binding site and a reverse transcription template (RTT). The design of the pegRNA has... we performed high-throughput ‘self-targeting’ screens with oligo libraries where pegRNAs are coupled ...cell contexts. Figure 1: High-throughput screening of pegRNA efficiency and building prediction tools...sequence-based prediction models from self-targeting screens such as PRIDICT/PRIDICT2.0 provide valuable insights...your target. Figure 3: High-throughput screening of the effect of chromatin on prime editing efficiency...

  18. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    Published
    Feb. 4, 2016, 3:30 p.m.
    ...plasmid, how will it be sent to me? Description: This describes how the plasmid or other product (kit...useful to me? Plasmid purpose: This is a brief description of what the plasmid was designed and used for...vector’s plasmid number here. Gene/insert: This describes what has been inserted into the backbone, and ...they’ll end up on the N or C terminus of the transcribed and translated insert. If it is an empty vector... sequencing. Growth in Bacteria: This section describes how to grow the bacterial strain in which the ...

  19. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    Published
    Sept. 9, 2014, 8:20 p.m.
    ...Unlike promoters which will create unique mRNA transcripts for each gene that is expressed, multicistronic...resistance gene. These reporters allow you to easily screen or select for cells that are expressing your gene...interest at high levels. Unlike vectors that express screenable or selectable markers from a unique promoter,...your gene as they are both derived from the same transcript. Of course multicistronic vectors do not have...efficient translation of multiple genes from a single transcript. The two strategies most widely incorporated ...incorporated into plasmids for research purposes are described below. IRES Elements Translation in eukaryotes usually...

  20. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    Published
    May 19, 2016, 2:30 p.m.
    ...for transcriptional activation from the 5’ LTR) and Rev (facilitates nuclear export of transcripts). A...Enable Genome-Scale, Knockout Screening Read about Genome-Wide Screening Using CRISPR/Cas9 Additional...both for targeted modification and genome-wide screening.   Lentiviral vectors in the clinic Since lentiviral... vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nat Methods. 2014...

  21. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    Published
    Dec. 2, 2014, 7:23 p.m.
    ...functions such as epigenetic regulation and transcription by using locus-specific chromatin immunoprecipitation...worked extensively with enChIP systems. Can you describe this technology and its advantages? Fujii: In ...biochemical analysis of genome functions such as transcription and epigenetic regulation. To elucidate molecular...enChIP system using CRISPR technology. Can you describe this technology and some of its potential applications... any other technologies that you would like to describe?  Fujii: I believe that "Biology is Method", i.e...Hodaka Fujii lab's Addgene page and click the "Subscribe to Alerts" link in the Addgene Alerts box. ...

  22. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    Published
    May 15, 2014, 3:33 p.m.
    ...biochemical properties. In 1995, Roger Y. Tsien described an S65T point mutation that increased the fluorescence...backbones for constructing fluorescent fusions. Transcription Reporter: Placing GFP under the control of a...promoter in a given cell type. This type of transcription reporting was among the earliest uses of GFP... gene of interest, or expressed from the same transcript as the gene of interest, but after an internal... GFP are available. Others: We’ve really just scratched the surface of the potential applications for ...to identify particular cell populations in drug screens, to visualize micrometastases in nude mice in cancer...

  23. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    Published
    July 25, 2019, 12:03 p.m.
    ...expression method enables us to quickly screen and investigate transcription factor candidates and the regulation...technique that uses a virus and the plant's post-transcriptional machinery to silence genes of interest (Unver...use of introns, which will be removed from the transcripts in the plant but not from Agrobacterium. Test...transient leaf transformations are a great help in screening candidate genes and speeding up research. Certainly...Budak. "Virus-induced gene silencing, a post transcriptional gene silencing method." International journal...

  24. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...protease allows a transcription factor to enter the nucleus and modulate transcription of the mCitrine ...your studies.  Find the plasmids to set up your screening systems at Addgene! Schematic of the evolution...their BioRxiv post, the authors generated and described several improved genetically encoded fluorescent...Takayuki Iwaki, Kimiko Hara, and Kazuo Umemura have described a simple method which alleviates the inclusion...activating expression of TCF4 while monitoring transcript and protein levels. Plasmids for multiplexed ...

  25. Plasmids 101: Codon usage bias

    Type
    Blog Post
    Published
    Sept. 27, 2018, 1:09 p.m.
    ...create secondary structures in RNA transcripts Effects on transcription (remember, it’s not all about translation...locations or fail to make it through the entire transcript resulting in the production of nonfunctional ...every amino acid I’d like to encode,” but, as described above, not every region of a protein should necessarily...translation - e.g. codon choice may interrupt transcription factor binding sites) As you might imagine,...algorithms and DNA synthesis companies such as IDT and GenScript host online codon optimization tools. Keep in ...

  26. Tips for Writing a Good Cover Letter

    Type
    Blog Post
    Published
    May 21, 2024, 1:30 p.m.
    ...experience and skills would help you do the job as described. Let’s imagine you’re applying to a job that ... your cover letter, you can expand on that by describing how you developed and used a ddPCR assay to answer...start by looking at the requirements in the job description. For each requirement, ask yourself if you have...exactly what you would want to talk about! You could describe your organizational system, your rationale for...company/organization]. I am a [adjective that describes you] scientist with over [number] years of experience...the university]” Include keywords from the job description to help the hiring manager quickly make the connection...

  27. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    Published
    Nov. 10, 2016, 3:30 p.m.
    ...antibiotic’, very often we immediately imagine a prescription for a bacterial infection. While this post will...that the term ‘antibiotic’ can also be used to describe anti-parasitic drugs and anti-fungal drugs, because...development of resistance include (Figure 1): Over-prescription of antibiotics (for example, giving antibiotics...not finishing the whole course of antibiotics prescribed to them This may seem counter-intuitive considering...The take-home? Always take your medication as prescribed. How do we know which bacteria are resistant ...infection is still effectively treated. We can prescribe antibiotics for the shortest time period possible...

  28. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    Published
    June 3, 2021, 1:15 p.m.
    ... rounds of screening. Created with Biorender.com.   Some phage display libraries screen antibody genes...expression systems. scFvs are also small enough to be screened for with in vitro display methods such as phage...and VH sequences. Phage display also allows for screening scFv generated from the B cell antibody genes ...the ways antibodies are. scFvs are even used to screen for new antibodies. To do this, antibody variable...variable domain sequences are expressed as scFvs and screened with in vitro assays to select for strong binders...

  29. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    Published
    May 13, 2025, 1:15 p.m.
    ...They rely on a viral protein called reverse transcriptase to convert their RNA genomes into DNA to be ... larger protein: PR (protease), RT (reverse transcriptase), and IN (integrase). These viral packaging ...the retroviral life cycle for a more detailed description). Since the integrated portion does not contain... cycle initially proceeds as usual. Reverse transcription of the transgene occurs in the cytoplasm, then... McCray, 2010). However, IDLV genomes are transcriptionally active, leading to transient expression of...Lentiviral vectors tend to integrate into transcriptionally active regions of the genome (Schröder et ...

  30. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    Published
    April 4, 2024, 1:15 p.m.
    ...SSC-H, or FSC-W/SSC-W, respectively. H, A, and W describe the shape of a histogram, which graphically represents... of the cell’s illumination (see Figure 2). H describes the maximum signal strength, while W results from...photocurrent vs. time. The resulting histograms is described through maximum current (H), the time the cell...depends on cell type and laser voltage. The voltage describes an electric potential that can be applied to the...comparing two markers simultaneously, we commonly describe the resulting populations as double positive (...that there are three parameters — H, W, and A — describing the intensity and duration of the detected signal...

  31. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    Published
    Aug. 2, 2016, 2:30 p.m.
    ... are isolated from the resulting colonies, and screened for the desired modification. Finally, the positive...final circular plasmid product pictured here. B) Screening of recovered plasmids for the mutation of interest...mutation). Similar results can be obtained with PCR screening using the primers specified by the arrowheads ... vastly facilitates the subsequent process of screening for succesfully mutated clones. Forward and reverse...can also form colonies under these conditions. Screening If a restriction site was introduced (or ablated...ablated), bacterial colonies can be screened by identifying the presence (or absence) of that particular ...

  32. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    Published
    Oct. 12, 2023, 1:15 p.m.
    ... Table 1 gives a brief overview of published descriptions of commonly used or emerging capsids. Selected...2017). AAV-PHP.eB has frequently been used for screening candidate regulatory elements such as enhancers...type-specific promoters or Cre lines. Spatial transcriptomics suggests that AAV-PHP.eB, when paired with ...capsid of choice for future promoter/enhancer screening. However, AAV-PHP.AX packages viral genomes at...target gene expression (EGFP, in the depositing manuscript) across the brain. AAV-BI-28 has been tested ...when using a ubiquitous CAG promoter. Spatial transcriptomics shows that its tropism is relatively more biased... Gradinaru lab. It was selected from a library screened in marmosets. AAV.CAP-Mac effectively transduces...

  33. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

    Type
    Blog Post
    Published
    May 18, 2018, 7:32 p.m.
    ...from the event can be found below along with a description of the program beneath it. You can jump to the...the different time stamps in the description to watch any sections you’re particularly interested in, ...abbreviated methods section found within a published manuscript. Luckily, many organizations have been working...JoVe, and Bio-protocol all work to provide more descriptive and interactive protocols that make it much easier...would get in-principle acceptance of the final manuscript regardless of whether the ultimate results were...Minisymposium on Reproducibility Booklet - Including descriptions of many organizations promoting reproducibility...

  34. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    Published
    Sept. 29, 2016, 2:30 p.m.
    ...identify possible gene function. In this post, I will describe a useful and cost-effective method that our lab...and probe with live cells The following is a description of a technique published by Simpson, Sacher, ...be cloned into E. coli and subsequent proteins screened for cell-binding abilities (6). This can be accomplished...long-term library storage. After overnight growth, scrape up, pool and extract plasmid DNA directly from ...construction for functional genomic and metagenomic screening." Applied and environmental microbiology 74.5 ...Protein-Protein Interactions Use CRISPR For Genome-Wide Screens Resources on Addgene.org Visit Our Microbiology...

  35. What's New in CRISPR - September 2019

    Type
    Blog Post
    Published
    Sept. 10, 2019, 12:56 p.m.
    ... fluorescent proteins, fusion proteins, or transcriptional regulators. In this system, Cas9 generates ... Find the CasX plasmids Drug inducible transcriptional activation and genome editing Yu Wang's lab...developed drug inducible CRISPR/Cas9 systems for transcriptional activation and genome editing by fusing Cas9...library with SpdCas9 to perform pooled CRISPRi screens to identify essential genes and phage host factors...metastasis cohorts and a prior mouse metastasis CRISPR screen. The Chen Mouse Tumor Suppressor Gene CRISPR Knockout...

  36. 6 Steps to Submitting a Resume That Gets Seen

    Type
    Blog Post
    Published
    July 9, 2015, 2:30 p.m.
    ...experience. Most Addgene PhD level job descriptions clearly describe that the roles do not involve bench ...submission of a job application. 3) Read job descriptions carefully Be open minded and be bold. You don...qualifications to apply for a position.  A job description is usually a wish list and the hiring manager... way to a hiring manager. Learn to confidently describe your strengths in your resume and don't be shy...

  37. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    Published
    Nov. 2, 2021, 1:15 p.m.
    ...sequence prior to an ATG start codon to prevent transcription in cells where the recombinase was not present... transgene into the correct orientation for transcription. Upon recombination one of lox2272 sites will...expression. To test their hypothesis, they randomly scrambled one pair of lox sites and either used the same...transgene (homologous pairs) or used different scrambled sequences to flank the transgenes (non-homologous...amplification. On the left homologous pairs of scrambled cre-recombinase sites lead to greater frequency...leaky expression. On the right non-homologous scrambled recombination sights lead to reduced reversion...

  38. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    Published
    Aug. 5, 2025, 1:15 p.m.
    ...efficiency! Screening mutant libraries After generating your mutant libraries, you have to screen thousands...generating the library of mutants and how they are screened. When conducting antibody evolution experiments...across large mutant libraries, high-throughput screening techniques such as ELISA and cell surface display... on the surface of cells. This enables direct screening via flow cytometry for improved binding under ... for Protein Engineering: library generation, screening, and affinity maturation. Methods in Molecular...

  39. Scientific Peer-review: Providing Critical and Kind Feedback and Advocating for Open Science

    Type
    Blog Post
    Published
    Aug. 8, 2019, 1:19 p.m.
    ...the beginning of my review - "The manuscript by ZZZ et al., describes ... The authors share a wealth of... you can ask that data and resources described in the manuscript are accessible to the community, increasing...recognize the hard work that the authors put in the manuscript. As a non-native English speaker, I sometimes...

  40. Revamp Your Lab Meetings With Creative Virtual Collaboration

    Type
    Blog Post
    Published
    Feb. 9, 2021, 2:15 p.m.
    ...make the screen-sharing experience more interactive. The button is on the right of the Share Screen control...control panel on the top of the screen and every participant get to annotate things as they come up in their...their minds. The final screen can also be saved and exported as a picture. I believe that showing something...pivotal in writing publications/grants too. As described by several works (Wuchty et al., 2007, Vicens ...

  41. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    Published
    June 8, 2017, 2:30 p.m.
    ...of Amsterdam. The previous post in this series described a practical approach to selecting a bright fluorescent...literature are performed in different ways. We describe a number of issues with some of the experimental...fluorescence, which, in its simplest form, can be described by a mono-exponential decay (analogous to radioactive...). Consequently, the intensity decay is often described by the t½, which is the time after which half ... al, 2008; Bindels et al, 2017) and cannot be described by a single parameter. Therefore, it is necessary...

  42. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    Published
    July 24, 2018, 12:51 p.m.
    ...CRISPR pooled libraries that allow functional screening of thousands of genes in genome-wide experiments...dividing and non-dividing cells. More detailed descriptions of the lentiviral elements and the 2nd and 3rd...regulation, gene of interest expression, knockout, and screening. There are many more useful viral vectors available... lab. Learn more about CRISPR plasmids for transcriptional inhibition. Retroviral Plasmid Kit Retroviral...collection can be used for in vivo phenotypic screening of small-molecule libraries and can be combined...

  43. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    Published
    April 6, 2017, 1:02 p.m.
    ...or gene copy. The coding sequence for GFP is transcribed and translated along with the gene, producing... last exon of the gene to ensure it would be transcribed and translated. For an N-terminally tagged protein...each cell line. The editing strategy and design described above can be used to create similar donor plasmids...Endogenous gene tagging with fluorescent proteins as described here enables live cell imaging to study cellular...

  44. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    Published
    Oct. 12, 2017, 2:26 p.m.
    ...user wants to carry out a high-throughput genetic screen, it is expensive and time consuming to individually...each insertion. Also, it remains challenging to screen for the genetically modified strains that contain...simultaneously introduce a selection marker for screening modified strains. With SapTrap, the user first...selectable marker (floxed Cbr-unc-119) for easy screening of the insertion event, and a variety of connector...hundreds of genes, generating powerful genetic screening libraries. Additional protein tagging systems ...

  45. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    Published
    Dec. 1, 2016, 3:30 p.m.
    ...template with the endogenous gene. Here, we'll describe the techniques and the plasmids used to inactivate... a rare event, so you must select as opposed to screen for cells where recombination has occurred. Neomycin...Conditional knockouts The methods and plasmids described in the preceding sections are simple ways of knocking...knockout, researchers often use the Cre/Lox system described earlier. In this case, you design your targeting... this gene will still be expressed. You'd first screen for this specific recombination outcome using PCR...

  46. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ...nickase system is based on the Cas9 D10A nickase described in Figure 4 of the Cong et al. (2013) Science ... The human U6 promoter prefers a 'G' at the transcription start site to have high expression, so adding...co-transfect in HR donor. Alternatively, a new system described recently is the double nickase system, with PX335...destroy the BbsI sites, a double digest should discriminate between positive and negative clones. Clones..., how can cells with that mutation be selected/screened? A13: Before starting your experiment, consider...sequencing, for example) on each colony in order to screen for positive ones.  -- OR -- If your HR template...

  47. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

    Type
    Blog Post
    Published
    April 2, 2019, 1:44 p.m.
    ...you can use to study these genes! Groucho, a transcriptional co-repressor The first conference room, that...Addgenies, was named after the protein Groucho, a transcriptional co-repressor important in development. Groucho.... It binds to a specific set of DNA-binding transcription factors to repress the expression of genes primarily...Ish-Horowicz. "The Groucho/TLE/Grg family of transcriptional co-repressors." Genome biology 9.1 (2008): ... Jiang, Jianqiao, and Helen White-Cooper. "Transcriptional activation in Drosophila spermatogenesis involves...

  48. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    Published
    Nov. 19, 2020, 2:15 p.m.
    ...If you’re new to Rett Syndrome, this blog post describes the history of Rett Syndrome research, the role...MECP2 in Rett Syndrome Rett Syndrome was first described in 1966 by Austrian physician, Andreas Rett, who...maturation (Tillotson & Bird, 2019) and is a global transcriptional regulator of thousands of genes. Studies have...have suggested roles in transcriptional repression, activity dependent de-repression, chromatin remodeling...groove of AT-rich DNA sequences to regulate transcription (Baker et al., 2013). Mutations that affect ...

  49. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...in vitro and in vivo. Incorporating reverse transcriptase (RT) variants, PE6a and PE6b are 500–800 bp ...LOV-Turbo plasmids here! Next, Qin and Cheah et al. described TransitID, where two proximity labeling enzymes...recently created a system for recording the transcriptional history of a cell. Lin et al. generated a fusion... HaloTag dyes and showed they could record transcriptional activation events as bands of induced GFP-iPAK4...Analyzing the ticker tape images could recover transcriptional histories, accurate to within 40 minutes. ...

  50. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    Published
    July 15, 2025, 1:15 p.m.
    ...developed a separate script for manually inputting known innocuous differences. This script compares the sequence...disregards the specified discrepancies. Any sequences that are a perfect match from this script can then continue...time, but does not leave much flexibility for discrepancies in backbone sequences. Most theoretical sequences...
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