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We narrowed to 793 results for: lat

Showing: 421 - 440 of 793 results

  1. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    Published
    July 14, 2016, 2:30 p.m.
    ...barely fits into AAV when you add in a gRNA and regulatory elements. To solve this problem, the Zhang lab...throughout the 23-base sequence. Double mismatches ablated Cpf1 activity, except when they were present in...future edits. In contrast, since Cpf1 cleaves relatively far away from the PAM, NHEJ might retain the ...mechanism. To prevent new editing post-HDR, repair templates should remove the PAM sequence. Many research ...

  2. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    Published
    April 28, 2016, 2:30 p.m.
    ...genetic tool for researchers to conditionally manipulate gene expression in vivo using site-specific recombination...orientation, the SSR will excise the DNA fragment. By manipulating the number, orientation, and type of target ...morphology, and circuit mapping of a targeted neuronal population.    Figure 2 (modified from Schnutgen...study the physiology or behavior of certain cell populations.  Acknowledgement:Through a partnership with...

  3. Gaining Leadership Skills Volunteering at a Professional Organization

    Type
    Blog Post
    Published
    Aug. 6, 2015, 2:30 p.m.
    ...where I gained invaluable experience working on and later chairing the Advocacy Committee. I’m now furthering...more. Having met many people at that conference, I later became involved with the NPA beyond the meeting....more productive and makes it easier to build relationships. Nonetheless, the committees would not function..., I had to learn how to develop new working relationships with these individuals who often had different...

  4. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    Published
    Feb. 7, 2019, 2:16 p.m.
    ...each gene, we needed to design guide RNAs, donor templates and sequencing primers for quality control. At... taken up to a month to design a single 96-well plate for HDR that satisfy all requirements. To alleviate...guides by CFD score, and it mutates HDR donor templates to avoid re-cutting when needed. It gives you ...the standard information that goes into a 96-well plate and comes out of it. Biotech companies are known...

  5. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    Published
    July 12, 2018, 1:09 p.m.
    ...Thursday, or, at the latest on Friday. It’s best to split cells before the plates are more than 80-85%...Benjamin Franklin)! It’s good practice to label your plates and/or flasks with your name, date, the cell line...disorganization, apoptosis, and gene expression disregulation on HeLa cells." Journal of applied phycology27.6...

  6. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    Published
    June 22, 2017, 2:30 p.m.
    ...protein or its role in a disease, researchers often isolate proteins of interest and examine them using biochemical...protein, it's important to validate its function relative to the untagged, wild-type protein. You may, for...the protein of interest and then analyzing the relative colocalization of their fluorescent signals and...One should also be aware of additional problems related to the properties of FPs themselves. These can ...

  7. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    Published
    Feb. 4, 2025, 2:15 p.m.
    ... need to deliver the Cas protein altogether. Platforming AAV Interest in interrogating and mapping the...glioblastoma, and generate programmable screening platforms for mouse tissues. Your AAVancement could be next...highly scalable cell type-specificin vivoscreening platform. bioRxiv (Cold Spring Harbor Laboratory). https..., Kalamakis, G., Lischetti, U., Jabaudon, D., & Platt, R. J. (2023). Transcriptional linkage analysis ...

  8. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    Published
    Jan. 30, 2024, 2:15 p.m.
    ...heartbeat – a muscle contraction – is directly related to the electrical impulse of the cardiac cells....propagated through multiple cells to do things like translate perceived signals to physical actions. Understanding...serotype specificity during delivery (more on this later). Chemigenetic sensors take a hybrid approach and...JF). The voltage-dependent changes in rhodopsin modulate the degree of quenching of the fluor via FRET....

  9. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    Published
    June 25, 2024, 1:15 p.m.
    ...2024). Chromatin context-dependent regulation and epigenetic manipulation of prime editing. Cell. https:/...primer binding site and a reverse transcription template (RTT). The design of the pegRNA has been shown...editing for precise and highly versatile genome manipulation. Nature Reviews Genetics, 24(3), Article 3. ...

  10. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

    Type
    Blog Post
    Published
    Oct. 30, 2018, 1:04 p.m.
    ...of a 5-6 day old human blastocyst [6]. They are isolated mostly from abnormal and/or supernumerary preimplantation...Characteristic ESC ASC Self-renewal High Low once isolated from the human body Usage Universal. Can generate...concern High. Embryos are destroyed during ESC isolation. Low Chances of post-transplantation rejection...when used therapeutically High Low (if ASCs are isolated from the patient) For many years, ESCs or ASCs...

  11. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    Published
    Oct. 17, 2017, 12:55 p.m.
    ...communicate the nuances of protein structure in a flat 2-D images. In the early days of structural biology...full-color images of proteins. But they are still flat. To truly appreciate the 3D structure of these proteins...   He then said “I hadn’t realized how perfectly flat the complex is on this surface”….where the complex...cleavage”. Jacob computationally added the non-template strand to the Jinek structure so that the resulting...

  12. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    Published
    Aug. 4, 2016, 2:30 p.m.
    ...institutions recently reported their inability to recapitulate the results of Gao et al 2016 in a letter to...NgAgo DNA guide is inexpensive and can be 5’-phosphorylated in house using a simple PNK reaction or can... the instability of the supposedly alien 5’-phosphorylated ssDNA guides, which might be rapidly degraded... an ssDNA guide. It is possible that, during translation, only a small fraction of nascent NgAgo protein...

  13. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ...double-stranded break. HDR copies the sequence from a repair template with flanking sequence homology for error-free...systems, and NHEJ will be favored even if a repair template is present. HDR also presents a cloning dilemma...CRISPR gene editing. In organisms where HDR is downregulated, MMEJ represents another method for making targeted...’s become clear that the power of this editing platform lies in the diversity of nucleases and their applications...

  14. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    Published
    Sept. 23, 2014, 3:05 p.m.
    ... up after trying out the new protocols.io beta platform for sharing, annotating, and storing life science...molecular cloning protocols, including making LB agar plates, restriction digest of plasmid DNA, and bacterial...bacterial transformation. And more plasmid-related educational resources can be found in our Plasmids 101 blog...the repository. These protocols show up on the related plasmid pages and can help scientists use these...

  15. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    Published
    May 29, 2014, 1:29 p.m.
    ...blastn), Standard Protein BLAST (blastp), and Translated BLAST (blastx). NCBI has a terrific getting-started...gaps in the nucleotide sequence, you can run a Translated BLAST (blastx) search to check for expected ORFs..., due to the additional searches involved in translating the nucleotide sequence into all six possible...choose for your sequencing result and can easily translate it, we recommend using blastp over blastx. The...

  16. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Published
    July 31, 2018, 12:55 p.m.
    ...field to a grinding stop and only now, 2 decades later, is it finally back in the limelight. On December...and severe visual impairment that faithfully recapitulated the human condition. Luck brought all these ...X-linked retinitis pigmentosa, achromatopsia, and age-related macular degeneration are ongoing and some will ...capsid variants, designing expression cassettes to regulate and optimize transgene levels, and large scale...

  17. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    Published
    July 31, 2020, 12:30 p.m.
    ...would convert adenine to inosine, which the translational machinery treats like guanine. This RNA editor...would permit point mutations in RNA which could recapitulate or rescue known pathogenic alleles, or introduce...ADAR will preferentially edit an adenine if the template has a cytosine mismatch at that position. PspCas13b-ADAR2DD...RNA Editing Capabilities through RESCUE Two years later, the Zhang lab expanded the RNA editing toolkit ...

  18. The Importance of a Fun Workplace: Company Culture at Addgene

    Type
    Blog Post
    Published
    Sept. 3, 2015, 2:30 p.m.
    ...trivia, and everything in between). Eleven years later,  Addgene is a company of fifty instead of three...situation- do we work so hard so we have time to play later? Or, does our play encourage us to work hard? Whatever...employees. The teambuilding culture helps us build relationships and thereby work better together.I realize I...encourage everyone to make an effort to build relationships with their colleagues. It can be easy to get...

  19. Using AAV for Neuronal Tracing

    Type
    Blog Post
    Published
    Aug. 9, 2018, 1:04 p.m.
    ...bi-directionality. For predominately anterograde tracing, biotinylated dextran amines (Veenman et al., 1992) or fluorescent...., 2007; plasmids at Addgene). AAV vectors: A platform for delivering neuronal tracers AAV vectors have...overview see Addgene’s AAV guide. AAV particles are relatively small (20 nm) in size which makes them capable...Transport: Cargo-Specific Mechanisms of Motility and Regulation. Neuron 84(2), 292–309. PubMed PMID: 25374356...Veenman, C.L., Reiner, A., and Honig, M.G. (1992). Biotinylated dextran as an anterograde tracer for single-...

  20. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    Published
    Jan. 17, 2019, 1:38 p.m.
    ... added on a regular basis, so the ecosystem of related tools is always growing. Beyond the ability to ...formatting. In this way data is moved from a static and isolated vehicle and becomes part of the dynamic and connected...understand and/or attempt to reproduce research relating to data deposited into a repository access to ...can be viewed – and conceivably additional work relating to reproducing the original research can be documented...
Showing: 421 - 440 of 793 results