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We narrowed to 863 results for: TOR;

Showing: 261 - 280 of 863 results

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

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...the activator - the CRY2 photolyase homology region (CRY2PHR) fused to a transcriptional activator domain...directed by the guide RNA (gRNA), while the CRY2PHR-activator fusion floats freely, depicted in the figure below...and CIB1 proteins heterodimerize and move the activator into position to activate gene transcription. ...manner. With their light-activated CRISPR/Cas9 effector (LACE) system, the Gersbach lab utilized a similar...fusion of full-length CRY2 and the transcriptional activator domain VP64. Using this system in HEK293T cells...photoswitchable proteins derived from the fungal photoreceptor, Vivid (VVD, N. crassa) (Kawano et al., 2015...transcription, they fused BldD to the transcriptional activators p65 and VP64 which in turn transcribes N-Cas9...

  2. DAP arrays for multiplex gene editing

    Type
    Blog Post
    Published
    July 7, 2022, 1:15 p.m.
    ... and progenitor cells. Now with the DAP array, it is possible to use the state-of-art base editor, prime...prime editor, or any CRISPR-based gene editors, for correcting mutations or installing functional genes...thought to enable MBE by using Cas12a fused base editors (dCas12a-BE). However, the poor editing efficiencies...they found led them back towards Cas9 fused base editors (nCas9-BE), as Cas9 is mechanistically more favored...number of gRNAs that can be packaged into viral vectors for potential in vivo applications or multiplex...array.7 Using DAP arrays with a variety of base editors, the Gao Lab successfully demonstrated up to 31...unprecedented number of genes with a single viral vector.   Fig. 1: Schematic of multiplex gene ...

  3. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ...adjacent fragment or vector. 5′ overhangs are generated on all the fragments and the vector, specifying the... together through transformation, so the cloned vector must be ligated. But in LIC, longer single-stranded...due to the absence of the other dNTPs. Once the vector and insert are (separately) digested to create ...received an important update, courtesy of Addgene depositor Stephen Elledge. His new method, named sequence...homologous recombination. SLIC inserts and linearized vectors must have homologous DNA sequences (20–60 bp) on...complete plasmid. Besides being compatible with any vector, the SLIC protocol itself is flexible. For one ...digestion (no dNTPs needed), allows both insert and vector to be digested in the same tube. Alternatively,...

  4. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    Published
    Oct. 5, 2021, 1:15 p.m.
    ...calcium indicator (Shemetov et al., 2021). In addition to FRET, intensiometric calcium indicators NIR-GECOs...proteins (NIR FPs) were derived from natural photoreceptors incorporating biliverdin IXα (BV) autocatalytically... derived from bacterial phytochrome (BphPs) photoreceptors have higher apparent cellular brightness than...were derived from cyanobacteriochrome (CBCR) photoreceptors (Oliinyk et al., 2019). They were engineered...performance, and applications of near-infrared calcium indicators in this recent review (Shcherbakova et al., 2021...encoded near-infrared fluorescent calcium ion indicator. Nat Methods 16, 171-174 (2019). Matlashov, M..... A near-infrared genetically encoded calcium indicator for in vivo imaging. Nat Biotechnol 39, 368-377...

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

    Type
    Blog Post
    Published
    April 22, 2015, 2:06 p.m.
    ...genes. We’ve created enough regulatory elements, CRISPRs, repressors, activators. We’ve been putting together...CRISPR/Cas genetic circuits are linked to the regulatory networks within E. coli host cells. ...in building genetic circuits is getting enough regulatory proteins that don’t interfere with each other...programmable that you can make a very large number of regulators. That basically means larger genetic circuits...that could conceivably be as large as natural regulatory networks. Addgene: What kinds of things could... pattern, shape. All of that is controlled by regulatory networks in the plant. If we want to access that...CRISPR is how much easier it is to work with than regulatory proteins. There are still things we haven’t solved...

  6. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    Published
    July 28, 2016, 2:30 p.m.
    ...massive amount of information regarding which regulatory proteins, signaling molecules, mutations, and...little is known about the order or timing of these factors. Recombinase-based state machines (RSMs), which...In the absence of extra recombinase-specific co-factors (which should be excluded from the system), these...paper (1). Lastly, it is possible to build gene regulatory RSMs (GRSMs, the subset of RSMs that regulate...also demonstrates that genetic parts (genes, terminators, and promoters) may be interweaved onto a GRSM...parts used in this paper (recognition sites, terminators, promoters, and genes) were built with flanking...them. We recommend using strong, directional terminators and promoters with buffer sequences on their ...

  7. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    Published
    July 11, 2019, 1:26 p.m.
    ...Mice can be engineered by inserting a targeting vector containing the gene of interest into a zygote, ...microinjection of DNA or infection with a viral vector containing the gene construct can be used to insert...generated via homologous recombination. A targeting vector is electroporated into embryonic stem cells and...engineer knockin mice. Co-injection of a targeting vector containing homologous ends to the breakpoint promotes...promotes homologous recombination of the targeting vector. CRISPR/Cas9-based systems have been developed ...the expression of a particular gene. Targeting vectors used to create knockout mice include a reporter...standing technique is the injection of a targeting vector into embryonic stem cells, which are then injected...

  8. Developing Transferable Skills During Science Training

    Type
    Blog Post
    Published
    June 3, 2014, 2:14 p.m.
    ...Looking for a mentor or want to start a peer mentoring group? "What Makes a Good Mentor?" Form Your Own...form a peer mentoring group to get real feedback Write whenever you can, find supportive editors so your ... Own Peer Mentoring Group: A How-To Guide for Scientists Download our eBook: Mentoring for Scientists ...your PhD or perhaps you have almost completed a postdoctoral position… or two. You have learned a lot. Whether... scientists, create and stick to timelines Be a mentor in a formal program Start something (a journal,... plan a conference Recruit many supporters and mentors that will speak for you (and you for them) Communication...

  9. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    Published
    Sept. 24, 2024, 1:15 p.m.
    ...the tool, and develop new applications for magnetoresponsive proteins. Motivated by curiosity Although...Maria dreamed about the possibilities of a magnetoresponsive fluorescent protein. Controlling an optogenetic...experiments to see if any small molecules act as a cofactor (among others, flavins are very effective) and...self-labeling FlavinTag made a reliable one-component magnetoresponsive system that could be expressed and purified...thrilled to see that MagLOV’s “astonishing” magnetoresponse was clearly visible whether expressed in E....or in purified form, and didn’t need any added cofactors.      Figure 2: Fluorescence intensity...respond to magnetic fields?” and “What are magnetoresponsive fluorescent proteins good for?”, and while...

  10. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    Published
    Dec. 1, 2016, 3:30 p.m.
    ...: A knockout targeting vector designed to insert a resistance gene. The vector contains a neomycin resistance... homologous sequence using a specially designed vector that contains sequence homology to the locus of... of the targeted gene have been cloned into the vector on either side of an antibiotic resistance gene...Figure 2). Figure 2: A knockout targeting vector designed to insert and later remove a resistance... the sequence to be inserted is cloned into the vector between the homologous sequences together with ... Figure 3: A knockout/knock-in targeting vector designed to insert GFP into a given locus. ...earlier. In this case, you design your targeting vector such that a set of three LoxP sites flank the resistance...

  11. Tips to Make the Most of a Scientific Conference

    Type
    Blog Post
    Published
    May 23, 2019, 12:26 p.m.
    ...following: the elevator pitch, talks, the poster session, networking, and social media. The elevator pitch Before...research, gathered up your data into an exciting story, and are ready to present your findings at a conference... Before the conference Practice your “elevator pitch,” the 30 - 60 second blurb you will use to introduce...work you are familiar with. You can tweak your elevator pitch slightly if you know a bit more about the...or talk. After the conference After using your elevator pitch several times at the conference, you might...might notice some trends. Were there parts of your elevator pitch where the other person seemed confused, ...questions. If you or your lab identifies a potential collaborator from the meeting, reach out to them, pointing...

  12. Easing the Protein Purification Process with pCri

    Type
    Blog Post
    Published
    June 19, 2015, 3:08 p.m.
    ...Bacillus subtilis and the yeast Pichia pastoris. The improved secretory capacities of these microbes make ...this information, you can then pick an expression vector and expression system. If you find that your protein...not fold well, you should screen a few different vectors to see which gives you the most functional protein...coli: purifying from Bacillus subtilis and Pichia pastoris pCri also includes plasmids designed for protein...producing difficult-to-fold proteins. In addition, P. pastoris has the capacity to glycosylate proteins and may...peptide (SP) that routes the protein through the secretory pathway to improve folding and disulfide bond ...coli, the use of a thioredoxin A (TRX)-containing vector can improve disulfide bond formation. pCri also...

  13. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    Published
    Oct. 8, 2020, 1:15 p.m.
    ...include the laboratory personnel, the serum, the cell culture media, water baths, incubators, etc. Contamination...another. Multiple cell lines should not be stored in a single incubator since it increases the risk of cross-contamination... Addgene's viral vectors articles Find tips and tricks for working with viral vectors Resources on Addgene.org...Contamination! Tissue culture media often have indicators included in the media that turns yellow from ...proliferating shortly. Once they are present in the laboratory environment, they are difficult to eradicate....using expired products or reagents that have been stored for a longer time. Use fresh media as much as possible...tighten the flask cap before placing them in the incubators. This will also solve the problem of contamination...

  14. Hot Plasmids - October 2022

    Type
    Blog Post
    Published
    Oct. 6, 2022, 1:15 p.m.
    ... voltage indicator: JEDI-2P  by: Brian O'Neill  The François St-Pierre lab and collaborators have successfully...found an indicator that is optimized for 2-photon imaging of voltages. This includes monitoring of fast... plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. Think we missed...what you'll find in this post: Improved voltage indicator: JEDI-2P Novel class of light-gated potassium...successfully developed an improved voltage indicator (GEVI) called JEDI-2P. Using a custom, high-throughput... available as Cre-dependent, EF1ɑ-driven viral vectors (AAV1) - and you can also find several other constructs...screen for improved genetically-encoded voltage indicators (GEVIs) based on the ASAP sensors. The resultant...

  15. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    Published
    July 27, 2021, 1:15 p.m.
    ...petunia SL enzymatic receptor DAD2. SLs induce a conformational change of the receptor which modulates the...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...inducible CRISPR inhibition (CRISPRi). A) Lentiviral vector design. B) Strategy for leak-free transcriptional...detection of SLs. The authors also showed that the receptor-based fluorescent biosensor has a high sensitivity...biosensor based on the parasitic Striga hermonthica SL receptor HTL7. This novel type of biosensors could be used...  Design and mechanism of action of the receptor-based SL fluorescent biosensor. Image from Chesterfield...CRISPR plasmids are always being added to the repository. To find all of the CRISPR plasmids available...

  16. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    Published
    Sept. 16, 2020, 1 p.m.
    ... to deliver larger cargo such as base editors and prime editors. While using AAV to deliver large transgenes...problems with Cas9 delivery. Adeno-associated viral vectors (AAV) are commonly used for in vivo gene delivery...SpCas9) and a gRNA together (~4.2 kb) into an AAV vector is challenging due to its packaging capacity of...feasible, it leaves little room for additional regulatory elements. Feng Zhang's group previously packaged...packaged Cas9 and multiple gRNAs into separate AAV vectors, increasing overall packaging capacity but necessitating...for Cas9. However, to allow for more room for regulatory sequences while still fitting Cas9 and its gRNAs...Split inteins have also been used to express base editors. References Chew WL, Tabebordbar M, Cheng JKW...

  17. Mesothelioma - Causes, Symptoms, and Treatment

    Type
    Blog Post
    Published
    Dec. 13, 2016, 3:30 p.m.
    ...including the lung, the heart, and the abdomen. Long story short, when asbestos is breathed in (or less frequently... for years, or they are exposed even one time. Doctors and researchers don’t entirely understand why some...do, but we do know that there are some genetic factors involved. For example, multiple studies show that... being done to see what, if any, other genetic factors may be in play. However, there is no question that...asbestos exposure remains the single-most important factor in developing mesothelioma. Asbestos, and certain...Early detection, immunotherapy, and angiogenesis inhibitors New research is also going into two other areas...exposed to asbestos Tests like these will allow doctors not only to detect mesothelioma much earlier, thus...

  18. CRISPR 101 eBook...en español

    Type
    Blog Post
    Published
    Jan. 18, 2024, 2:15 p.m.
    ... Daniel Mejías Sánchez, PhD, traductor  L. Denise Roche, PhD, MD, editora, [email protected] The ...with distributors across the world to help increase global access to materials in our repository. This...  Daniel Mejías Sánchez, PhD, translator  Denise Roche, PhD, MD, editor, [email protected] la traducción, Addgene contrató a una traductora científica profesional, L. Denise Roche, para...el acceso global a los materiales de nuestro repositorio. Este increíble regalo permitirá que más personas...Addgene contracted with a professional scientific translator, L. Denise Roche, to edit the translation. We...

  19. Celebrate National Book Month with Addgene

    Type
    Blog Post
    Published
    Oct. 3, 2019, 12:47 p.m.
    ...skeptical reader of historical fiction (I can personally attest to this!). Following history from the Ice Age...relevant science."     - Randall Carter, Contracts Administrator Inferior: How Science Got Women Wrong - and ...and the New Research That's Rewriting the Story by Angela Saini "This book is a smart and carefully presented...science of race."     - Joanne Kamens, Executive Director  I Contain Multitudes by Ed Yong "Microbes are... Tsang, Science Communications and Marketing Coordinator Non-science favorites  The Mistborn series by...another secret."     - Marcy Patrick, Associate Director of Quality Control Jonathan Strange & Mr. Norrell...Norrell by Susanna Clarke "A quasi-historical/fantasy fiction. Two dueling magicians in the 1800s in England...

  20. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    Published
    May 20, 2021, 1:15 p.m.
    ...you can usually see a director sitting in their chair not watching their actors live on set, but peering...about getting in close to mic them up. Monitor: I then set up a monitor connected to the wireless transmitter...camera, OR, monitor from that distance.  Build: We got a wheeled stand and mounted a monitor to it. It ... their mask. But while the camera is here, the operator is in a different room.   Design: The previous...peering at a 7-inch wireless monitor. Wireless video technology has gotten very impressive, but due to costs...the beating heart of Addgene. It’s why we exist, storing plasmids, and distributing them to labs that need...Cycle 5 - The Future! Figure 5: The mobile monitor and teleprompter being repurposed for a different...
Showing: 261 - 280 of 863 results