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We narrowed to 849 results for: HA

Showing: 161 - 200 of 849 results

  1. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

    Type
    Blog Post
    ...It’s clear that CRISPR-Cas9 technology has really changed the game for anyone looking to quickly and ...human genes. Unlike the GeCKO library, each library has been enriched for different gene targets. These pooled...system. Genome wide mouse CRISPR library The Yusa lab has leveraged the CRISPR system to target all 19,150 ...stably transfected for Cas9 expression. The Yusa lab has used their library to screen for genes involved in...

  2. The time and cost required to make a plasmid

    Type
    Blog Post
    ...five years, inflation has occurred. Here at Addgene, the consumables cost per stab has increased 50% since...make a plasmid. Doench’s lab in the Broad Institute has created hundreds of thousands of plasmids, using ...since 2019, while overall inflation of USD has gone up 19.8% in the same time period. For ease of calculations...per plasmid, Addgene, since its founding in 2004, has saved the scientific community a collective and very...

  3. Sharing is Caring: But How to Distribute Open Hardware?

    Type
    Blog Post
    ...sharing open hardware has a similar goal. The sharing of modules, ideas, and blueprints has the potential not...Since its early years, the light microscope has been a powerful tool in almost all disciplines. In biology...Sharing open-source plans that rely on such tools has allowed the community to regularly replicate designs...portable device for studying plankton using flow cells, has become an established alternative to high-priced ...

  4. Working with Nuclear Receptors

    Type
    Blog Post
    ... funding model in nuclear receptor research, and has generated a set of comprehensive datasets that collectively...interactome of these key regulatory molecules, and has provided clues to their previously uncharacterized...website. NURSA and Addgene The great progress that has been made in the NR field over the last thirty years...and coregulators, and although the reporter gene has over the years evolved from chloramphenicol acetyltransferase...research tools is a double-edged sword, however, and has greatly increased the burden associated with identifying...

  5. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ... the best outcomes. The go-to in vivo option AAV has become the preferred delivery method for in vivo ... other viral vector delivery methods. Lentivirus has limited packaging options, with most lentiviral vectors...possibilities when designing screens. Each serotype has its own tropism, or a preference to infect specific...BioRender.com. Beyond the identified serotypes, AAV also has psuedotypes. Psuedotyping is the process of making...Lentiviruses have capacities of 8–12 kb, while AAV only has approximately 4.7 kb — which means that when using...the genetic landscape of tissue-specific diseases has grown exponentially, aided by advancements in in ...

  6. Addgene’s Top 10 Blog Posts from 2014

    Type
    Blog Post
    ...repository. Our blog is just over a year old, and it has grown steadily during 2014. We were excited to have...continues to grow as the genome editing technique has developed and improved. The CRISPR posts written ...about. Our post on “Making Your Own Competent Cells” has been extremely popular, generating many additional...

  7. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ... applications (v82 has higher affinity for glutamate but slower kinetics; v857 has faster kinetics but...iGluSnFR3 (Aggarwal et al. 2022). This new version has higher signal to noise ratio and is better localized...for two-photon microscopy. The resulting JEDI-2P has faster response kinetics and increased brightness...and deep-tissue imaging. Vladislav Verkhusha’s lab has generated a new genetically encoded calcium indicator...interest, it’s important to choose a biosensor that has a binding affinity close to the range of concentrations...physiology and metabolism under various conditions. LiLac has improved response size and reduced sensitivity to...

  8. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ... but inadequate access to reagents and equipment has become a bottleneck. In the last few years, scientists...nucleic acids at the atomolar level. The Zhang lab has adapted natural RNase activity of the Cas13 protein...Gootenberg et al., 2017 and 2018). While the Doudna lab has used Cas12a’s non-specific ssDNA degradation to develop...field (Myhrvold et al., 2018). Now, the Zhang lab has shared a protocol for using SHERLOCK to detect SARS-CoV...at atomolar levels, within one hour. DETECTR thus has the ability to rapidly detect nucleic acids with ...diagnostic applications. The company Mammoth Biosciences has been started based on DETECTR technology with the...Biotechnology. Like the SHERLOCK detection system, DETECTR has not been approved for diagnostics yet, but is undergoing...

  9. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...specific protein folding needs. The pCri system has recently been used to study a matrix metalloproteinase... Queensland’s Institute for Molecular Bioscience has provided a solution for this problem by developing...eukaryotic cell extracts (Gagoski et al 2015). He has also constructed a library of eGFP-tagged human ORF...constructs. The Cell-Free Protein Expression Kit has been used to produced recombinant proteins for in-vitro...populations through time, Christopher Marx's lab has engineered and validated a new method called FREQ-Seq...cost-effective manner with minimal bias. The FREQ Seq kit has been used in microbiology studies monitoring sequence...binds your RNA sequence of interest. The Zhao lab has made this easy with their PUF Assembly Kit. The kit...

  10. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ...excision site after cleavage by the transposase. It has a cargo capacity >100 kB, although integration efficiency...efficiency decreases with cargo size. Sleeping Beauty has a close-to-random integration profile in mammalian...hypersensitivity sites. Like Sleeping Beauty, piggyBac has an integration efficiency in human cells similar ...1996). Unlike Sleeping Beauty and piggyBac, Tol2 has a weak consensus sequence for its preferred target...hypersensitivity sites. Tol2 is only active in vertebrates and has lower efficiency of integration in human cells than... is a truncated version of the original Tol2 and has ~3-fold increase in transposition activity  Tol2...encodes a Type I-F CRISPR-Cas system. This system has four major components: 1) a CRISPR RNA (crRNA), 2...

  11. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...nervous system (CNS) (Wu, Asokan, et al., 2006). AAV1 has also been found to transduce skeletal muscle, cardiac... the most common choice for AAV pseudotyping. It has broad tissue tropism, including liver, muscle, lung...generally the best choice for targeting astroglia. It has additionally been reported to have tropism for lung...through our in-stock viral prep service.  AAV8 AAV8 has high transduction efficiency in multiple tissues,...applications targeting liver tissue. In addition, it has been shown to transduce skeletal muscle, cardiac,...through our in-stock viral prep service.  AAV9 AAV9 has relatively broad tissue tropism and high transduction... the CNS (Issa et al., 2023). Additionally, AAV9 has been shown to transduce liver, skeletal muscle, pancreatic...

  12. Important Considerations When Using AAVs

    Type
    Blog Post
    ...graduate program at the University of Manitoba. Our lab has been utilizing chemogenetics (Designer Receptors ...Although AAV2 can transduce many tissue types, it has low expression in several brain regions where other...serotypes to infect different cell types. In the CNS, it has been shown that there is variable transduction of...from the Original Location - Axonal Transport It has been demonstrated that certain serotypes can undergo... of movement, distance traveled, etc.) (4). This has also been identified with AAV2 (5), AAV6 (6) and ...required for the onset of gene expression. However, it has yet to be determined what gene expression looks like...problem in many experiments, a group of researchers has identified this feature of AAVs as a new tool for...

  13. The Future of Research Symposium Boston 2015

    Type
    Blog Post
    ...Research will be held from 22-24 October. This blog has been contributed by guest blogger and Future of Research... of a system that, despite all of its successes, has many deep-rooted flaws (1). As we discovered through...example, Harvard’s Therapeutics Graduate Program has a compulsory internship to be taken in an industry...Marder, professor of Biology at Brandeis University, has recently published an article arguing against reducing...Labor and Worklife Program at Harvard Law School. He has continually promoted the cause of increasing data...

  14. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    ...methods. Each method has its own pluses and minuses, but Golden Gate cloning has been especially useful...number of potential clones. Golden Gate assembly has a few advantages over other cloning methods. Exonuclease-based...just a few steps. More recently, CRISPR technology has adapted Golden Gate cloning for inserting the appropriate...also be adapted to express multiple gRNAs. Addgene has two Golden Gate-based gRNA assembly methods available...

  15. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    ...THE ORIGINAL NgAgo ARTICLE DISCUSSED IN THIS POST HAS BEEN RETRACTED AND FOLLOW UP STUDIES HAVE FAILED ...Natronobacterium gregoryi, or NgAgo for short. Addgene has already distributed this plasmid all over the world...Publication at Addgene Cas9 and NgAgo head to head NgAgo has a number of unique features compared to Cas9. These...compare CRISPR and NgAgo, but it seems that NgAgo has lower potential for off-target effects. In Gao et...

  16. Addgene's Top 10 Blog Posts of 2015

    Type
    Blog Post
    ...2015 has been an exciting year for the Addgene blog. We’ve released over 80 posts on a diverse array ...Scientists and the Plasmids 101 2nd ed, the 3rd edition has now been released), and have started providing recorded...incredibly happy to see that one of our career posts has made it to the top of the list this year. While we...doing just that. Do you like the direction the blog has taken over the past year? Do you think there are ...

  17. What's New in CRISPR - Winter 2018

    Type
    Blog Post
    ...dual-nuclease Cas9-Cas9 chimeras Scot Wolfe’s lab has developed dual-nuclease Cas9-Cas9 chimeras with higher...been many Cas nucleases developed for plants, it has been difficult to compare the performance of different...recently discovered in 2017. Now Dominik Niopek’s lab has adapted anti-CRISPR proteins for optogenetic control...

  18. A Scientist's Guide to Ebola

    Type
    Blog Post
    ... disease once limited to small pockets in Africa has spread to urban areas and across the ocean. A nurse... considered unlikely to change – but the illness has been fatal in 70 percent of the most recent cases... in the New York Times and on his blog, where he has offered to answer readers' questions. If you have...

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

    Type
    Blog Post
    ... and software development has caught up with it. The peer review process has been challenged with many...targetable NGG PAM sites whereas the human genome has about 300 million. With such a sparsely targetable...needs of each individual; the  community as a whole has yet to settle on any true kind of value. I’ve dubbed... genome editing tool, the CRISPR/Cas9 technology has been surrounded by a whirlwind of research activity...significant improvement. As a result, the software piñata has been filled with slight variations on a theme without...evaluate off-targeting, but currently no software has been designed for this purpose. This second area ...findings to help everyone else. CRISPR technology has reached into so many different communities that it...

  20. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...certain experimental setups. The brightness of NLuc has caught the attention of researchers wanting to perform...proteins can be excited by Nluc emission and the lab has dubbed these fusions LumiFluor Reporters. The LumiFluor...mNeptune2 through 33 mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness and improved maturation...with Luciferases can overcome these problems but has been hampered by dim signals from the coupled luciferase...ReNL; plasmid #85203) NLuc fusions. The Nagai lab has additionally deposited more than 25 fusions of the...Pairing NanoLuc® luciferase with fluorescent proteins has allowed researchers to make innovative tools not ... Technical Services Scientist, Kyle Hooper, PhD, has also worked with R&D for product development and ...

  21. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    ...foot shock that has become associated with a visual or auditory cue. The hippocampus has long been known...every other advanced technique in neuroscience it has a bit of a steep learning curve and many opportunities...within your own neuroscience sub-discipline that has published using the technique and ask if they would...sophistication of mouse transgenics. Everything that has been described is intended for the mouse; however... behavioral models. Beyond rodents, optogenetics has even been applied to other species such as non-human...

  22. Bringing Sustainable Practices to the Lab: Easy Action Items

    Type
    Blog Post
    ...marketing campaign! MIT Green Labs has a lot of resources about this, and even has a list of biological samples...are stored long term at -70 °C. My Green Lab also has resources and even a freezer challenge that your ...

  23. Download Science-Themed Zoom Backgrounds from Addgene

    Type
    Blog Post
    ... backgrounds all year and want a change? Addgene has you covered. Today we’re releasing 11 bold science-themed...fan of Addgene or our very cute mascots!  Blugene has been helping us make Plasmids 101 videos and needed...background.           Maybe your viral research has taken you in the direction of COVID-19. We have a...

  24. Your Top Requested Plasmid in 2015!

    Type
    Blog Post
    ...by Neville Sanjana. This fantastic research tool has been requested from the repository nearly 1,000 times...provided by the Zhang lab. Even though lentiCRISPR v2 has only been available for a little over a year and ...and a half, a small sampling of the research it has already been used for includes work probing mTOR signaling...

  25. Inntags: Innovative Protein Epitope Tagging

    Type
    Blog Post
    ...need new epitope tags? Short epitope tags like FLAG, HA, and Myc can adopt a number of different conformations...effects of GFP fusions to inntags or traditional FLAG, HA, and Myc tags. Impressively, IT6-GFP did not reduce...observed with IT5 and IT6 fusions. In contrast, Flag and HA tags had especially detrimental effects on cell size...

  26. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...avoid selecting for cells in which the construct has randomly integrated into the genome, a negative selection... opposed to screen for cells where recombination has occurred. Neomycin, puromycin, and hygromycin resistance...still functions normally because one of its exons has simply been replaced with the same sequence flanked...flanked by LoxP sites while the resistance cassette has been placed into an intron.    Figure 4: ...second the floxed exon.   After recombination has occurred, you'll first remove the resistance marker...

  27. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    ... in NCBI’s numerous databases. As our repository has grown over the years (we now have over 60,000 plasmids...we analyze as part of our quality control process has steadily grown. On a busy week, we may need to analyze...our quality control process. Consequently our team has refined our use of the BLAST web browser interface...also enjoy seeing how our quality control process has changed with the introduction of next generation ...BLAST (blastp), and Translated BLAST (blastx). NCBI has a terrific getting-started guide for BLAST, which...

  28. Identifying Sequence Elements with SnapGene's Feature Database

    Type
    Blog Post
    ... makes software good? Fortunately, that question has been thoroughly answered by experts in human-computer...specialized controls available when needed. SnapGene has been engineered to be easy and enjoyable to use. ...protein sequence match even when the DNA sequence has changed. Identifying plasmid control elements Coding... with these extensive efforts, such an algorithm has limitations. For instance, it could miss a common...

  29. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    ...Transcriptional Programs with CRISPR RNA Scaffolds." Adam has also created a video to help scientists visualize...various flux imbalances. The conventional approach has typically been to either systematically delete/overexpress...introduce heterologous genes, but the trend of research has shifted in recent years toward tinkering with regulatory.... Since the technique relies on base pairing, it has promise to be host-independent, beyond the oft published...

  31. Technical Design of a Western Blot

    Type
    Blog Post
    ...agent than BME and has a less… distinctive smell. BME smells like rotting eggs but has a longer shelf life... It doesn’t degrade during long-term storage and has both high binding capacity and high protein retention... specialty low-background PVDF membrane, it also has higher background noise than nitrocellulose membranes... other hand, doesn’t require any pre-wetting and has lower background than PVDF. It is a little quicker...quicker to use, with high binding capacity, though it has lower retention after binding and washes. It’s less...and need to be read fairly soon after the reaction has started. Types of chemiluminescence enzymes The two...darkroom and a developer. Film is highly sensitive but has a limited dynamic range, which means it may take ...

  32. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    ...Cas9-adenosine deaminase fusion Caixia Gao’s lab has developed a new plant base editor from an evolved...in diverse bacteria from various environments. It has the advantage of stable genomic integration and is...Cre or Flp recombinases used to excise markers. It has the advantage that CRISPR efficiency can be controlled...

  34. Making Career Development an Integral Part of the Science Training System

    Type
    Blog Post
    ...My son is an engineer and my daughter has a business degree. From the very start of their undergraduate...reduced impact of scientists and science. The pandemic has demonstrated that, more than ever, we need all of...at University of Massachusetts Medical School. It has been supported by a Steering Committee and working...

  35. Designer PUF Proteins for Any RNA Target

    Type
    Blog Post
    ...– to specific sequences of DNA has become commonplace. This ability has opened up a world of possibilities...the original paper says). In their hands, BsaI-HF has produced less reliable results. She also notes that...

  36. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    ...specific field(s). Most anyone who has spent any time in academia has likely had the opportunity to attend... one that no one sees a purpose in attending and has lost both its focus and critical mass of participants...their own knowledge of the field. Perhaps interest has simply waned. When attendance slumps below a certain...rebooted or disbanded all together once their purpose has been served. I hope that with some of the information...

  37. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...Biotech in May 2016 (1), NgAgo has received much attention and the plasmid has been requested by labs around...THE ORIGINAL NgAgo ARTICLE DISCUSSED IN THIS POST HAS BEEN RETRACTED AND FOLLOW UP STUDIES HAVE FAILED ...spotlight and take a moment to remember that NgAgo has only been available for genome editing for a few ... synthesis, which could be less desirable if one has to target multiple sites in a single experiment. ...

  38. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    ...popularity. A yellow flame indicates that a plasmid has been ordered more than 20 times; red, more than 50...available through Addgene. Publication: If the plasmid has been published, the paper in which it was first used... their new plasmid.  The Addgene Vector Database has more information about common vector backbones.  ...plasmid number here. Gene/insert: This describes what has been inserted into the backbone, and provides information...gene or DNA element of interest. If the depositor has indicated that there is a particular mutation found...

  39. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...Counter-selection with sacB has revolutionized allelic exchange. Even now, only half our lab has switched over to..., 2014). In recent years, CRISPR-Cas9 technology has also aided in the creation of scarless mutations,...isolated are those where the allelic exchange vector has undergone single crossover and integrated into the...to pTOX. However, we feel it has clear advantages, and are hopeful that continued development of allelic...the lab of Matthew Waldor at Harvard. Now that he has an efficient allelic exchange system, he is so excited...

  40. Five Popular Model Organisms

    Type
    Blog Post
    ...more commonly known: the fruit fly. The fruit fly has been used in scientific research for over a century... a transgenic mouse line. Over the years the fly has become an ideal model organism to study an array ...development, genetics, and the nervous system. The fly has also emerged as an important human disease model ... described in 1993 by Norbert Perrimon’s lab and has been continuously improved upon since then. If you...Zebrafish (Danio rerio) Since the 1960s, the zebrafish has gradually become an important model organism. They...Caenorhabditis elegans) The worm, Caenorhabditis elegans, has been used extensively as a model organism since the...
Showing: 161 - 200 of 849 results