Skip to main content
Addgene
Open Menu

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 784 results for: SCR;

Showing: 251 - 300 of 784 results

  2. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    ...plasmid with its elements described below.   Vector Element Description Origin of Replication ...for further study. Promoter Region Drives transcription of the target gene. Vital component for expression...

  3. New CRISPR Web Resources and #12DaysofCRISPR Recap

    Type
    Blog Post
    ...certainly taken notice! Our newest page in the Transcriptional Regulation category already has five blue flame...with eSpCas9, SpCas9-HF1, & HypaCas9 Day 10: Transcriptional regulation: Truncated gRNAs for Regulating ...Chavez) Day 11: Pooled libraries: Genome-Wide Screening Using CRISPR/Cas9 and Plasmids 101: NGS Quality...

  4. Plasmids 101: Positive and Negative Selection for Plasmid Cloning

    Type
    Blog Post
    ...positive selection, negative selection, and/or screening. We’re focusing on positive and negative selection...in this blog post, but don’t worry, we’ll cover screens in a future post. Positive selection: Cells that...with or without your inserted DNA) and further screening will help you determine if the plasmid backbone...

  5. Addgene at the ASM Microbe 2016 Conference

    Type
    Blog Post
    ...well-optimized CRISPR screening technology. Addgene got several shout-outs for making popular screening libraries ...John Doench’s opening presentation at the CRISPR screens symposia outlined the fantastic work this Broad...

  6. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    ...CRISPR/Cas inspired genome editing was initially described by Cong et al. and Mali et al. in January 2013...Nine months later, more than 1500 publications describe work to improve the tool’s specificity, orthogonality...orthogonality, and multiplexibility in various species and describe an assortment of applications. Despite the ubiquity...

  7. Viral Vectors 101: What is a Virus (and a Viral Vector)?

    Type
    Blog Post
    ...retroviruses, reverse transcribe their gRNA into DNA, which is then transcribed into mRNA before being...depending on the class of virus. RNA genomes can be transcribed into mRNA, directly translated into proteins,...

  8. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    ...(WHP) Posttranscriptional Regulatory Element (WPRE) forms a tertiary structure when transcribed which ... of RNA and RNA translation, and promotes RNA transcript longevity. The polyA is located at the end of...

  9. Science Careers: Unruly Interests Feed Many Paths

    Type
    Blog Post
    ...programmed regulation of DNA replication and gene transcription during vertebrate embryogenesis. My interests...each day brings in new and surprisingly diverse manuscripts. I work with people, nurturing networks, as much...Career Decisions Check Out Our Career Resources Subscribe to Career Blog Posts Resources at Addgene.org...

  10. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    ...large-scale functional screen in bacteria. Sensor variants were then refined by a secondary screen in rat hippocampal...developed and published in 2010 for visualizing the transcription factor cAMP-responsive element-binding protein...

  11. DAP arrays for multiplex gene editing

    Type
    Blog Post
    ..., lengthy genetic payloads and insufficient transcription on the array have limited the scalability and...potential in vivo applications or multiplex CRISPR screening.   Using tRNAs Luckily, a fun feature of tRNA...engineering of crops or allow for sophisticated CRISPR screenings that would perturb the unprecedented number of...

  12. Early Career Researcher Toolbox: Free Tools for Science Writing

    Type
    Blog Post
    ..., it's hard to get those words out and onto the screen and transform your work into a paper or a grant... categorized either based on the section of a manuscript they might be found, i.e. introduction, methods...the Addgene blog: Tips for writing scientific manuscripts: literature searching, reading, and organizing...

  13. Virtual Science Conference Coming Up? Three Addgenies Share Their Experience

    Type
    Blog Post
    ...to keep the audience engaged through a computer screen. For the Reproducibility for Everyone Workshop,...author of the poster,” he says.Viewing a poster on a screen is challenging, requiring you to zoom in to different... their poster, eliminating the need to zoom and scroll around the poster (Reshef et al., 2020).The Twitter...

  14. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ...adoption of the technology. In 2012, Saito et al. described the first Nano-lantern, a fusion of a mutagenized...hours! The latest enhanced Nano-lanterns (eNL) described in Suzuki et al. improve the brightness of previous...substrate furimazine. In addition to the previously described applications, the enhanced signal of eNL allows...

  15. Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection

    Type
    Blog Post
    ...27478040. The lung cancer mutant collection is described in this paper. Systematic Functional Interrogation...the genetic pathway reference collections are described in this paper Systematic morphological profiling...  The genetic pathway reference collection is described in this publication We also wrote an extensive...

  16. Multiplex Genome Editing with CRISPR-Cpf1

    Type
    Blog Post
    ...editing!  A recent paper from Feng Zhang's lab describes how to use Cpf1 for multiplex genome editing.  ...transfection gRNAs expressed from a polycistronic transcript  Requires co-transfection of Cas9 Nissim et alTsai...transfection gRNAs expressed from a polycistronic transcript Requires co-transfection of Cas9   Xie et al  ...

  17. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    ...expressing cell lines, which can then be used both for screens (using pooled CRISPR knockout libraries) and individual...monoclonal Cas9-expression line prior to performing a screen or even a single gene knockout. Top AAV: pAAV-hSyn-DIO-hM4D... vectors and genome-wide libraries for CRISPR screening. Nat Methods. 2014 Aug;11(8):783-4. PubMed PMID...

  18. Kiran Musunuru on the Newest TALEN Genome-Editing System

    Type
    Blog Post
    ...Cell, Musunuru, Chad Cowan and their colleagues describe a much more efficient tool for doing that disease...modeling work in human pluripotent stem cells: a transcription activator-like effector nuclease (TALEN) kit...people unfamiliar with this new TALEN kit, can you describe it for us? Musunuru: TALENs are a type of genome...

  19. Summer SciComm Series: Cognitive Load

    Type
    Blog Post
    ...formulation of new knowledge than it is to read a manuscript whose whole purpose is to provide detailed evidence...images, noises and music, spoken words, and any onscreen text. Be aware of the effort the audience is expending...Maybe I needed to employ a visual aid instead of describing something verbally. Sometimes a group was just...

  20. CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt

    Type
    Blog Post
    ...Systems Biology in November, Voigt and Alex Nielsen describe what’s possible when multi-input CRISPR/Cas genetic...many deposits to Addgene over the years. Can you describe the toolbox you are putting together?  Voigt: ... Those will allow you to set the amount of transcriptional and translational activity a cell has and control...

  22. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...neurons from each class to fully characterize the transcriptome. The first data release for CeNGEN is planned... The 3’UTRs of mRNA are important for post-transcriptional regulation in C. elegans. To study the role...Addgene In addition to the wealth of open resources described above, there were many plasmid tools presented...

  23. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...'ve overlooked regulatory fragments of these transcripts. I remember when small RNA was just something...’t give tRNAs much thought. High throughput transcriptome sequencing methods including work done by the...circRNA datasets and download the custom python scripts needed to discover circRNAs in RNA-seq data. As...

  24. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    ...histone modifications, which can influence the transcription of genes by RNA polymerase II, or by the addition...methylcytosine in CpG islands makes  genes less transcriptionally active. The role of epigenetics in studying...of tumor suppressor genes can repress their transcription and inactivate them. Global hypomethylation,...

  26. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    ...chromoproteins have a shorter history. In 1987, scientists described an unusual blue-colored protein from the jellyfish...methods. While techniques such as  blue-white screening require X-gal, an expensive exogenously-added ...chromoproteins don’t require other substrates for screening. Thus chromoproteins can be used as markers in...

  27. Plasmids 101: Protein Expression

    Type
    Blog Post
    ...synthesize a particular protein DNA must first be transcribed into messenger RNA (mRNA). mRNA can then be translated...template of choice and allowing these cells to transcribe, translate and modify your protein of interest...assembled in vitro using purified components of the transcription and translation machinery. These include ribosomes...

  28. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...transfer plasmid at 30 °C instead of 37 °C and then screen for ITR recombination with a SmaI or XmaI restriction...duplicated regions like the ITRs. Restriction digest screening for ITR deletion is still necessary. To save time...mL of my AAV transfer plasmid cultures and then screen with SmaI digests. I freeze the bacterial pellet...

  29. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    ...following cloning methodologies have been extensively described in other articles; so if you are interested in... to assemble up to 6 different fragments in a prescribed order. These fragments can be pooled in one tube...homology. You might also consider FastCloning, first described by Chaokun Li and colleagues. Here you simply ...

  30. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ...transduced cells whenever possible. This post will describe our workflow for viral genome sequencing (VGS)...identification, we developed custom open-access Python scripts that can perform a deeper analysis of the data....Check out our Viral Vectors Featured Topic Page Subscribe to Viral Vectors Blog Posts Learn How We Use NGS...

  31. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ...vivo. A common choice for conducting genome-wide screens using CRISPR. AAV transduction Only compatible...containing gRNA and Cas9 are used in in vitro transcription reactions to generate mature Cas9 mRNA and gRNA...complexes Purified Cas9 protein and in vitro transcribed gRNA are combined to form a Cas9-gRNA complex...

  32. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    ...video_player "embed_player" overrideable=False, type='scriptV4', hide_playlist=True, viral_sharing=False, embed_button...into two components: 𝜃x(t) and 𝜓z(t). 𝜃x(t) describes the exponential decay process we observed, while...appears to return to normal. The model fit these two discrete phases with an estimate of 62.3% for the COVID...

  33. Transferable Skills Guide: Identifying Your Transferable Skills

    Type
    Blog Post
    ...looking at your resume while they wrote the job description. You check off all the boxes - you have the desired... gaps in scientists' transferrable skills. In describing the limitations of the study, the authors noted...over email. Examine your current and past job descriptions. Scan through all the things for which you are...

  34. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    ...vector with two other independently regulated transcription units. Created with BioRender.com.   Planning...contain plasmids with undesired ligations, so always screen multiple colonies after each transformation. This...beginners or those performing complex assemblies should screen a minimum of 8–10 colonies. You can check colonies...

  35. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ...heterologously by bacteria inside the body. So we screened gas vesicles gene clusters from a diverse set ...expression heterogeneity required single cell screening to isolate a clonal cell line capable of producing...improvement in expression. Stabilizing the gvpA transcript with WPRE reduced the number of copies of gvpA...

  36. How to Write a Scientific Review Article

    Type
    Blog Post
    ...we probably exchanged dozens of drafts of the manuscript. This is the time to transform the document into...Resources on the Addgene Blog Writing Scientific Manuscripts: Literature Searching, Reading, & Organizing ...Organizing Tips for Improving Your Next Manuscript 6 Tips for Grant Writing Resources on Addgene.org Check out...

  37. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ...amplification) or reverse transcriptase (RT)-RPA, respectively. RPA is coupled with T7 transcription to convert amplified...2018) CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity. Science 360...

  38. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ...used in research, and data repositories. Below I describe how each of these kinds of connectivity facilitates...deposits will be made. The Dataverse integration is described in this video and the Figshare integration in ...from RSpace documents to external data sources (described above) further enhances the value the repository...

  39. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...restriction endonucleases in plasmid addition was first described by studies of EcoRI and Bsp6I in Escherichia coli...systems. As with the other toxin-antitoxin systems described above, one part of the pair is more stable than...toxin-antitoxin system discovered. Unlike the systems just described, the hok/sok systems consist of three components...

  40. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ...expected for postdocs in many cases should be as described in the notice of the new NRSA levels and in the...very relieved not to be going through the urgent scramble of their peer institutions. The issues facing ..., and it is becoming ever clearer that outside scrutiny of science and the academic system is increasing...

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

    Type
    Blog Post
    ...Cas13a as a diagnostic tool, as discussed below. Subscribe to CRISPR Blog Posts Applications of Cas13a   ...Recombinase Polymerase Amplification (RPA) or Reverse-transcription RPA (RT-RPA), respectively. Amplified nucleotides... the collateral RNA cleavage response is well described in bacterial cells, LwaCas13a does not display...

  42. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...methyltransferase — that can drive lasting transcriptional silencing (Nuñez et al., 2021). Now, in a collaboration...versions using dCas9, zinc finger proteins, and transcription activator-like effectors (TALEs), with fifteen...Weissman, J. S. (2021). Genome-wide programmable transcriptional memory by CRISPR-based epigenome editing. Cell...

  43. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...Easi-CRISPR in vitro transcribes RNA encoding your repair template, then uses reverse transcriptase to generate..., and ultraviolet light. In this post, we will describe the mechanism of homology directed repair, one...

  44. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ... The one-pot saturation mutagenesis technique described in Wrenbeck et al is a PCR-based approach for ...library that’s ready to be tested in a functional screen of your liking. This method is similar to site-directed... screen...

  45. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...strong transcriptional terminator BBa_B1002 (from IGEM) upstream of the MCS to reduce transcriptional readthrough...worked too. Rhamnose induction relies on the transcriptional activator rhaS. To allow manipulation of bacteria...

  46. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    ...same fluorescent markers, Brainbow 3.1 and 3.2 (described in the same publication) are unique because by...cell type. The Confetti mouse was first used to describe the self-renewal of cells in the intestinal crypt...previous Brainbow technologies were useful for discriminating neurons within only one field of view. Tetbow...

  47. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...site-specific incorporation of phosphoserine (as described below) provides researchers with a valuable tool...context-specific effects of UAG suppression, as described (6). To determine the stoichiometry of phospho-incorporated...phospho:non-phospho species on PhosTag gels as described above. Reagents and protocols Sep-OTSλ plasmid...

  48. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...applications, including transgenesis, gene-trap screens, and gene editing. How does the piggyBac™ system...effects, such as with signaling molecules or transcription factors, or when the gene is constitutively-..., since there is no selection, you will need to screen somewhere between one hundred and one thousand ...

  49. Antibodies 101: Validation

    Type
    Blog Post
    ...Antibody-independent methods may include analysis of transcriptomic or proteomic data from several different tissues... feasible and (2) large-scale proteomic and transcriptomic datasets already exist.   Example: You want...tissues. You know that there is a wealth of transcriptomic data from this sample type, so you use your...

  50. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ...brightness of individual fluorophores. Table 1 describes the brightness of a few commonly used fluorophores...brightness levels on your smartphone or computer screen), and you should be able to properly label many...panel! Designing a Panel In this example, I will describe a basic panel (Table 2, Figure 5) for a hypothetical...
Showing: 251 - 300 of 784 results