We narrowed to 255 results for: hal.2
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TypeBlog PostPublishedJuly 21, 2017, 12:48 p.m.... you can read more about them in review articles (2–4). You can also take a look at my recent paper to...Development 118, 401–15 (1993). PubMed PMID: 8223268. 2. J. B. Duffy, GAL4 System in Drosophila : A Fly Geneticist...two genes by one Gal4 driver will produce roughly half as much protein as overexpression of each gene individually...384–391 (2004). PubMed PMID: 15262411. 4. T. D. Southall, D. A. Elliott, A. H. Brand, The GAL4 System :...Marka, J. C. Kagan, Environmental Stress Causes Lethal Neuro-Trauma during Asymptomatic Viral Infections...
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Viral Vectors 101: Inverted Terminal Repeats
TypeBlog PostPublishedOct. 19, 2023, 1:15 p.m....ITR, which can either be “flip” or “flop” (Figure 2). Additionally, the ITR contains a 4-nucleotide Rep...site for Rep proteins (Savvy et al. 2013). Figure 2: AAV2 wild type ITR in flip and flop orientation. ...Packaging, and Potency of AAV Vectors. Microorganisms, 12(2), 310. https://doi.org/10.3390/microorganisms1202031038399714...10.1089/hgtb.2016.133. PMID: 28967288. Shitik, E. M., Shalik, I. K., & Yudkin, D. V. (2023). AAV- based vector... -
Targeted Mutagenesis with EvolvR
TypeBlog PostPublishedFeb. 21, 2019, 1:15 p.m....while increasing the standard mutation rate by only ~2-fold over nCas9 levels. Two sets of modifications ...10-3 mutations per nucleotide per generation, and (2) the addition of the thioredoxin-binding domain (TBD...components of EvolvR and its potential applications. Challenges of existing mutagenesis methods Several continuous...drawbacks with these approaches that makes it challenging for users to achieve targeted diversification...PMID: 30253789. PubMed Central PMCID: PMC6154929. Halperin, Shakked O., et al. "CRISPR-guided DNA polymerases... -
Click Biology: How to Click and Play to Explore Biology
TypeBlog PostPublishedOct. 21, 2025, 1:15 p.m.... engineered components, like protein tags (Figure 2). Through the work of many groups, there are systems... in Addgene's repository (Table 1). Figure 2: Adding function to My Favorite Protein (MFP) through...Exchangeable HaloTag Ligands HaloTag Reversible binding of fluorescent ligands to HaloTag PCV-Cas9, Cas9...Plasmid Collection HaloTag Collection of research tools from Promega, including many HaloTag plasmids Exchangeable... tube or in cells. There are also SNAP-tag and HaloTag modules, which can connect proteins to any kind...SNAP-tags or CLIP-tags Tetbow SNAP-tag, CLIP-tag, HaloTag Based on the Brainbow system, adds Tetracycline... -
Treating Muscular Dystrophy with CRISPR Gene Editing
TypeBlog PostPublishedJan. 26, 2016, 3:30 p.m....nonsense-mediated decay. Although Nelson et al. observed only 2% genome editing in one experiment, they found the ... cells throughout the body, especially stem cells 2. Remove any immunogenicity of the AAV vector Once ...Science. 2015 Dec 31. doi:10.1126/science.aae0169 2. Long C, Amoasii L, Mireault AA, McAnally JR, Li H...Precision genome editing faces the challenges above and more. One chief challenge is upregulating HDR in mature...complex oligonucleotide treatment comes with many challenges, researchers have begun to explore genome editing... the time to clinical approval. Even with the challenges precision editing faces, it’s encouraging to ...WX, Scott DA, Gootenberg JS, Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang... -
CRISPR 101: Epigenetics and Editing the Epigenome
TypeBlog PostPublishedJune 24, 2020, 5:45 p.m....form or as ready-to-use lentivirus. Figure 2: dCas9-Tet1 demethylates cytosines at promoters and...pX330a dCas9-LSD1 for targeted removal of H3K4me1/2 and H3K9me2 histone modifications. Like the TALE-LSD1...approach for targeted DNA demethylation. Cell Discovery 2: . https://doi.org/10.1038/celldisc.2016.9 Find plasmids... demethylase to demethylate enhancer regions (Mendenhall et al., 2013). By comparing gene activation when...sorting and selection of transduced cells. Grant Challen’s lab also created constitutive (pCMV-dCas9-D3A)...contributed to updating this article. References Haldeman JM, Conway AE, Arlotto ME, et al (2018) Creation...Fowler T, Rees R, Kramer A, Martens A, Edwards JR, Challen GA (2016) Reprogrammable CRISPR/Cas9-based system... -
CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More
TypeBlog PostPublishedSept. 28, 2020, noon... pCRISPomyces-2 contains Cas9 and a gRNA cassette. The simpler system of pCRISPomyces-2 displays a higher...and a subsequent curing protocol that requires only 2-3 hours incubation. E. coli and T. citrea scarless...four major components including (1) a CRISPR RNA, (2) four proteins forming the QCascade DNA-targeting ...DSB) are lethal. Addgene depositor Luciano Marraffini’s lab took advantage of this lethality to design...organisms, CRISPR-induced double stranded breaks are lethal because the non-homologous end-joining (NHEJ) repair...Cas9 will cleave the gene of interest, inducing a lethal DSB. This system is distinct from those used...While the activation of transcription is a more challenging problem than repression, scientists have deposited... -
Antibodies 101: Reading a Flow Plot
TypeBlog PostPublishedFeb. 6, 2024, 3:13 p.m....A-B+ cells. If you look at the dot plot in Figure 2, you might assume that all cell populations are positive... the different populations. In the case of Figure 2, one population is B- and another two are B+. Our ...are in the lower right corner. Figure 2: This flow plot has two populations that are B+ and...from flow cytometry experiments can be a little challenging to understand if you’ve never done any flow. ...., Sait, K., Anfinan, N., Al-Wasiyah, mohammad khalid, Huwait, E., Gari, M., & Al-Qahtani, M. (2018).... -
Quickest Way to Deposit Plasmids: The Deposit Spreadsheet
TypeBlog PostPublishedMarch 8, 2017, 3:15 p.m....our deposit spreadsheet to complete Step 1. Steps 2 and 3 are usually very easy — our tech transfer team...columns are shown in red in the spreadsheet (Figure 2) and are described in Table 1. Some cells have drop-down...Filling out and sending the deposit spreadsheet Figure 2: Example of the deposit spreadsheet showing the first... (budding yeast), S. pombe (fission yeast), A. thaliana (mustard weed), O. cuniculus (rabbit), synthetic... -
Pathways Over Time Plasmids Engage Students in Functional Genomics Research
TypeBlog PostPublishedMay 22, 2018, 1:38 p.m.... undergraduate students (1, 2), but educators face significant challenges in designing suitable projects... by western blotting in other experiments (Figure 2). Complementation reveals conservation of function...Biology Education: A Call to Action, Washington, DC. 2. President's Council of Advisors on Science and Technology... -
"What Makes a Good Mentor?" and 6 More FAQs About Science Mentoring
TypeBlog PostPublishedDec. 17, 2013, 2:47 p.m....fulfilling career in science can be challenging – is advice and guidance from only 2 people enough? For many years...portion of our careers. In that time we may have only 2-3 formal supervisors that will provide guidance and... -
Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies
TypeBlog PostPublishedNov. 27, 2019, 2:03 p.m.... characteristics of select AAV serotypes in Table 2 from the paper. Delivery of AAV into the brain can...routes for neuroscience see Table 1 from the paper. 2. Manipulate your cell of interest: Cell-type specific...development, learning, and behavior poses many challenges in neuroscience with many brain regions, neuronal... -
Plasmids 101: Degron Tags
TypeBlog PostPublishedMarch 14, 2024, 1:15 p.m....required components are 1) a SMASh-tagged protein and 2) asunaprevir (small molecule). dTAG The degradation...gone through an additional round of evolution (AID-2) to reduce some of the leakiness associated with the...Adapted from Nabet et al., (2018). HaloTag The HaloTag was initially developed by Promega as an ...the Halo-tagged protein in question, it is targeted to the proteasome for degradation. The HaloTag is ...straightforward system — HaloTag and its PROTAC are all that is needed to facilitate degradation. HaloTag is also the...visualization of proteins of interest. A fluorescent Halo ligand can be added to media, buffer, etc., which...proteolysis-targeting chimera (PROTAC) was developed for HaloTag. PROTACs are engineered molecules which target ... -
Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux
TypeBlog PostPublishedSept. 13, 2016, 2:30 p.m....is funded have been well documented over the past 2 years (Alberts et al, PNAS 2014; Taylor, eLife, 2016...PMID: 24733905. Pubmed Central PMCID: PMC4000813. 2. Taylor, Justin Q., et al. “Avoiding a lost generation...since the Future of Research Symposia Ongoing challenges facing the scientific enterprise and the way ...landscape of the scientific enterprise will provide challenges and opportunities for us all. For example, implementation...categorize all postdocs at institutions, will be challenging. At the same time, there is much opportunity ...the pressure on for further increases will be challenging, but will be aided by the development and implementation... is an important way to push the balance from challenge to opportunity. We hope to see you at the upcoming... -
Cellosaurus: A Cell Line Information Database
TypeBlog PostPublishedFeb. 21, 2023, 2:15 p.m....groups, such as their vaccine production, SARS-CoV-2 research, or adenovirus packaging cell lines dataset...-Line Knowledge Resource. J Biomol Tech. 2018; 29(2): 25-38....split you got from the graduate student down the hall who got it from some technician downstairs who got... -
CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems
TypeBlog PostPublishedJuly 23, 2020, 1:20 p.m....base pair to another base pair at a specific site. 2. Temporal, spatial, or conditional control of CRISPR...difficult-to-engineer Sulfolobus islandicus rod-shaped virus 2 (Mayo-Muñoz et al., 2018). Only viral particles that...Editing of Sulfolobus islandicus Rod-Shaped Virus 2. Viruses 10:695 . https://doi.org/10.3390/v10120695...that have the gene deletion could replicate when challenged with the S. islandicus CRISPR-Cas system. 5. ... -
Antibodies 101: Flow Compensation
TypeBlog PostPublishedMay 16, 2024, 1:15 p.m..... Let's look at an experimental example in Figure 2. Figure 2A shows the sample prior to compensation.... line with the other populations. Figure 2: Compensation example based on experimental sample..., you will see the populations move, as in Figure 2, following compensation. Figure 3: Flow plot...Compensating between your fluorescent markers can be challenging! But we hope this guide can help you get started... -
Antibodies 101: Single Chain Fragment Variables (scFvs)
TypeBlog PostPublishedJune 3, 2021, 1:15 p.m....panning and scFv characterization. Figure 2: Phage display starts with a pooled library of scFvs...the short epitope GCN4, also called a scaffold; and 2) a GCN4 binding scFv that’s fused to GFP. When the...Fv (Variable Domain) (scFv) Fragments. Antibodies 2:193–208 . https://doi.org/10.3390/antib2020193 Khantasup... the body or self antigens, and toxins that are lethal to animals. In the clinic, scFvs’ size also provides...plasmid in bacteria (Ahmed et al., 2012). One challenge of bacterial expression is the proper formation... -
Top Requested Lentivirus and AAV of 2016
TypeBlog PostPublishedJan. 6, 2017, 3:56 p.m.... tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One. 2013 ...PMID: 24086725. PubMed Central PMCID: PMC3785459. 2. Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher...makes generation of high titer Cas9 lentiviruses challenging. This cytotoxicity also causes growth rate variability...):337-47. PubMed PMID: 12595892. 4. Sanjana NE, Shalem O, Zhang F. Improved vectors and genome-wide libraries... -
Hot Plasmids: Summer 2025
TypeBlog PostPublishedAug. 28, 2025, 1:15 p.m....RNA guide (known as an omega RNA, or ωRNA) (Figure 2), but TnpB is small enough to be carried by viral ...than a third of the size of Cas9). Figure 2: Comparison of TnpB (A) and Cas9 (B) genome editors...Figure 5: Mechanism of ATLASCre. VAMP2 = synaptobrevin 2; At = ALFA-tag; BACE = β-secretase; AF = AMPA.FingR...perform neuronal tracing, anterograde tracing poses challenges, from non-selective anterograde transport to ...