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

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We narrowed to 402 results for: CAPS;

Showing: 121 - 160 of 402 results

  1. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...mutated to inactivate the protein’s nucleolytic capability, and this nuclease-null or “dead” Cas9 (dCas9...complementarity to a target site, Cas9’s DNA binding capability remains intact while its nucleolytic activity... RNA hairpin, such as the MS2 hairpin, that is capable of recruiting additional effector domains (4). ...with a ≤15nt gRNA that contains an RNA adapter capable of recruiting a transcriptional activator, robust...

  2. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...but only transiently. However new viral vectors capable of providing long-term gene expression in retinal...and severe visual impairment that faithfully recapitulated the human condition. Luck brought all these ...integration into the genome, and its large DNA packaging capacity is well suited for larger transgenes. Unfortunately...further improve the technology: engineering of new capsid variants, designing expression cassettes to regulate...

  3. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...affinity purification in situ of regulatory elements (CAPTURE) (Liu et al., 2017). This system utilizes dCas9...through proteomics and the chromosome conformation capture (3C) technique. In addition to 3xFLAG and biotin...when inappropriately expressed can cause facioscapulohumeral muscular dystrophy (FSHD), a disease for ...Central PMCID: PMC6286148. Liu, Xin, et al. "In situ capture of chromatin interactions by biotinylated dCas9...

  4. Technical Design of a Western Blot

    Type
    Blog Post
    ...binding capacity and high protein retention. It comes in a variety of pore sizes and protein capacity, allowing...are not with denaturing agents like DTT or β-mercaptoethanol.  Denaturing proteins To denature your proteins...you'll want to use SDS and either DTT or β-mercaptoethanol (BME). DTT is a stronger reducing agent than... is a little quicker to use, with high binding capacity, though it has lower retention after binding and...

  5. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ...look specifically at the characteristics and capabilities an ELN needs to facilitate true reproducibility...without access to them, the record of research captured in the ELN is incomplete. Thus, to support reproducibility...for discussion and review. It’s also possible to capture Slack conversations and import them into RSpace...and workflows that went into it. The ability to capture the conversations and associate them with a particular...

  6. The Twelve Days of CRISPR

    Type
    Blog Post
    .... If you missed it, here’s the recap! The twelve days of CRISPR recap On the first day of CRISPR, Addgene...day of CRISPR, Addgene gave to me: a method to capture promoter-enhancer 3D chromatin structures using...

  7. PEI Calculator for Planning AAV Packaging Transfections

    Type
    Blog Post
    ...Transfer Plasmid : 1 pHelper: 1 RepCap OR 3 Transfer Plasmid : 5 pHelper: 2 RepCap Select what type of tissue...using 28 μg of total plasmid (Transfer, helper, and RepCap added together) per 15 cm dish or 2500 μg of plasmids...

  8. Easing the Protein Purification Process with pCri

    Type
    Blog Post
    ...addition, P. pastoris has the capacity to glycosylate proteins and may recapitulate the post-translational modifications...yeast Pichia pastoris. The improved secretory capacities of these microbes make them useful for producing...

  10. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    ... can rupture E. coli spheroplasts to release encapsulated proteins. Video by ACS. For a more cartoony ...Harvard Medical School. Follow her on Twitter @jessicapolka.     References 1. Kanabrocki, J. A., Quackenbush...Protein Piston That Breaks Membranes to Release Encapsulated Cargo. ACS Synth. Biol. (2016). doi:10.1021/...

  11. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ... will be necessary to improve its multiplexing capacity. Multiplexing is key due to the natural redundancy...multiplexing in systems with limited packaging capacity. Still confused about the various tRNA expression...References Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system. Xie...

  13. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ... below:   ID Cv (Capsid) FLAG Tag Terminus 79629 (N) and 79628 (C) Cv (capsid) N and C 79633 (N...system has a much larger cargo capacity than lentivirus and is capable of efficiently creating stable ...homology (usually 30-40bp overlaps). To expand the capabilities of Gibson assembly and make a more modular platform...

  14. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    ...Recent developments promise to further change the landscape facing all scientists, both young and established...potential to profoundly shift the academic science landscape: the treatment of trainee scientists as workers...shifts caused to the practical and philosophical landscape of the scientific enterprise will provide challenges...

  15. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    ...counterparts, so they will be easier to deliver in low-capacity vectors like AAV. AAV (adeno-associated viral ...tissues. However, these vectors have a low genetic capacity of ~4.5 kb. At 4.1 kb, SpCas9 barely fits into...understand where Cpf1 fits into the genome editing landscape. References 1. Kim D, Kim J, Hur JK, Been KW...

  16. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    ...biology of the worm into a computer model that recapitulates how molecular biology impacts worm locomotion...mutant worm. Like many academic labs, we have the capability to study many different mutants because the Caenorhabditis...and cobble together quite a diverse array of capabilities. So don’t throw old equipment away until you...

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

    Type
    Blog Post
    ... to the establishment of in-house sequencing capabilities and Seqwell’s plexWell technology, we can now...small predetermined seed sequences unique to the capsid plasmid. Our recombination software looks for and... Moullier P, Salvetti A (2005) Evidence for Encapsidation of Prokaryotic Sequences during Recombinant ...

  18. An Introduction to Adenovirus

    Type
    Blog Post
    ... E1 region). The space in the viral particle’s capacity that is made available by this deletion from the... (and their structural proteins, which would encapsidate new virions) are not expressed. Note that replication-competent...genome, and adenoviral vectors all have a genomic capacity that is much higher than other viral vectors (...

  19. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ...transcriptase are packaged (Wu et al., 1997) inside of the capsid . This is useful for viral reproduction, and for...for reverse transcriptase (RT) is present in the capsid, as the engineered, recombinant genome is (thankfully...mouse embryo cell lines in which each cell has the capacity to be activated to produce infectious murine leukemia...

  20. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ...DNA transposons are ubiquitous genetic elements capable of spreading within and between genomes, and have...transposition. By natively combining the DNA integration capabilities of transposases and the target programmability...large DNA cargos into bacterial genomes. This new capability was named INTEGRATE (Insertion of Transposable...

  21. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    ...using electrophoresis, typically capillary electrophoresis. Capillary electrophoresis reads a fluorescent...M. B., Snyder, M. P., & Barron, A. E. (2011). Landscape of Next-Generation Sequencing Technologies. Analytical...

  22. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    ...AAV vector is challenging due to its packaging capacity of AAV (~4.7 kb). While this approach has been...separate AAV vectors, increasing overall packaging capacity but necessitating purification and co-infection...-AAV toolbox that retains the gene-targeting capabilities of full-length SpCas9. This set of plasmids ...

  23. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...that combines enrichment and imaging of proteins captured on magnetic beads (Arimura et al., 2024). In MagIC-cryo-EM...GFP fusion in this proof-of-concept study) are captured on streptavidin-coated magnetic beads through ...bundle; SAH: 60-nm single alpha helix) and target capture module (SpyTag-GFP nanobody) assembled on beads...

  24. Important Considerations When Using AAVs

    Type
    Blog Post
    ...interest. Although they have a small packaging capacity, AAV vectors were suitable for our needs for the...Joshua C., and Richard J. Samulski. "Packaging capacity of adeno-associated virus serotypes: impact of...recombinant adeno-associated viral vectors: packaging capacity and the role of rep proteins in vector purity....

  25. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...of interest as well as sequences necessary to encapsulate that insert into a viral particle (the packaging...cell culture as lentivirus-derived vectors are capable of transducing certain quiescent or terminally ...ELISA dosing method detects the expression of viral capsid antigen p24, the FACS dosing method correlates ...

  26. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ..., brighter, and specific dyes, expanding the capabilities of fluorescence-based techniques in biotechnology...Fluor HaloTag Ligands expand the self-labeling capabilities of this platform. The Janelia Fluor ligands .../doi.org/10.1007/s41048-017-0037-8 Lucidi, M., Capecchi, G., Visaggio, D., Gasperi, T., Parisi, M., Cincotti...

  27. Which Fluorescence Microscopy Technique is Best for Me?

    Type
    Blog Post
    ...entire field of view is excited at one time and captured in a single camera exposure. This is much faster...out-of-focus light. The most difficult characteristic to capture in these types of samples is their fast movements...imaging, the light microscope was limited in its capacity to resolve closely lying structures. This is because...

  28. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    ...sheath fluid and transported through tubes or capillaries to a laser. As the cells move through the fluidics...different directions. The degree of scatter is captured by detectors and provides useful information about...fluorochrome, emits a fluorescent signal that is captured by detectors. The strength of the emitted photons...

  29. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    ...to increase your output. But, exceeding a kit’s capacity can actually reduce your ultimate plasmid yield...ensure proper lysis, but you will still face column capacity limitations since the columns can be “clogged”...magnet as you scale up. Table 1: Standard Prep Kit Capacities* Prep size Amount of culture Approximate ...

  30. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    ...Cas9 fused to an engineered adenosine deaminase capable of chemically converting adenosine (A) to inosine.... They also developed a machine learning model capable of predicting editing outcomes of their various..., they added the pioneer factor SOX2, which is capable of binding to and remodeling inaccessible chromatin...

  31. Make a Splash: Notions of Scientific Impact Are Evolving

    Type
    Blog Post
    ...Ventures like Impact Story seek to find ways to capture scholarly work that exists outside of traditional...Of course, no one measure will ever adequately capture a notion as complex as “scientific impact.” As ...

  32. A Scientist's Guide to Ebola

    Type
    Blog Post
    ...It’s pretty hard to escape news and fears of Ebola these days, as the infectious disease once limited...described how the glycoprotein transforms itself to escape vesicles within host cells and turn the cell into...

  34. Reflections and Looking Ahead

    Type
    Blog Post
    ...scientific gains by adding long-read sequencing capabilities to complement our next-generation sequencing...flourishes when data are shared. The scientific landscape is evolving rapidly, and with it, the needs of...

  35. In Living Color: The Skinny on In Vivo Imaging Tools

    Type
    Blog Post
    ...described in Nature Communications last year, “The capability to control spectral properties of PA FPs with...is also looking in the direction of biosensors capable of revealing, for example, how muscles work inside...

  36. CASTing Off for New Shores in Human Genome Editing

    Type
    Blog Post
    ...power of CRISPR combined with the large sequence capacity of transposons open whole new worlds to explore... the course of evolution, Tn7-like transposons captured and repurposed nuclease-deficient CRISPR-Cas systems...

  39. Bricking Science: Portraying Scientific Reality Through LEGO

    Type
    Blog Post
    ...gnome. See this video where he/she/it/something is captured on camera: Unfortunately, unless they’ve worked...were mirrored by the nearly infinite regulatory capabilities of gene pathways just waiting to be discovered...

  40. Safe Port in a Storm...How Addgene is Weathering the Pandemic

    Type
    Blog Post
    .... We don’t raise money except occasionally for capital equipment or resources needed to launch a new service...workforce for when we need to be working at full capacity to serve the scientific community. First, we applied...
Showing: 121 - 160 of 402 results