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We narrowed to 303 results for: hal.2

Showing: 241 - 280 of 303 results

  1. Viral Vectors 101: The AAV Retrograde Serotype

    Type
    Blog Post
    ...axon terminals to the soma (retrograde tracer) (Fig. 2). Viruses as neuronal scalpels Viruses can be injected...recombinase-dependent vectors, this can cause challenges for the experimental design as these vectors ...

  2. Communicating Your Science With Help From ComSciCon

    Type
    Blog Post
    ...aspects of the work you presented may also be unclear. 2. Make the audience your primary focus When writing...of science, communication isn't easy.  But the challenge is worthwhile and rewarding. A truly passionate...

  3. AAV Q&A with Tim Miles

    Type
    Blog Post
    ...retransduce a model organism such as mice, e.g. first at 2 months, then at 4 months? Or do antibodies become ...prohibitive problem? A: Re-administration is a challenge for AAV due to the development of neutralizing...

  4. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    ...account when simulating a digest as depicted in Lane 2. Controlling methylation Finally, you can control...plasmids. It turns out that restriction enzymes are one half of naturally occurring restriction modification ...

  5. Optogenetics Guide

    Type
    Guide
    ...Cryptochrome 2 and CIB1 Blue-light–mediated induction of protein interactions based on Arabidopsis thaliana cryptochrome...OPTOSTIM PHR domain2 of cryptochrome 2 (Cry2) from Arabidopsis thaliana is fused to truncated forms of cytosolic...influx into the cell when stimulated. Channelrhodopsin-2 (ChR2), the first widely adopted optogenetic tool,...cryptochrome 2 and CIB1 450 LARIAT Inhibits protein function by reversibly sequestering targets into large...customizable TALE DNA-binding domain with cryptochrome 2 protein and its interacting partner CIB1 to control... with single-spike temporal resolution. PLoS One. 2(3):e299. PMID 17375185 Kim B, Lin MZ. 2013. Optobiology...comparative analysis of microbial opsins. Nat Methods. 9(2):159-72. PMID 22179551 Saunders A, Johnson CA, Sabatini...

  6. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...split at 175 Arabidopsis homolog of the yeast TREX-2 mRNA export complex: components and anchoring nucleoporin...Tebo et al. Nat Commun. 2019 Arnaud Gautier HaloTag TagBiFC allows exogenous labeling for single-molecule...and reversible complementation. TagBiFC (split HaloTag) provides maximum flexibility in color labeling...

  7. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...PMID: 24901218. PubMed Central PMCID: PMC4047077. 2. Pagliara, Patrizia, et al. "Ostreopsis cf. ovata ...Turning on the UV light 3-4 times a week for up to half an hour can help keep your hood contamination free...Additional Resources on the Addgene Blog The Challenges of Cell Culture Cloning Mammalian Cells with ...

  8. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    ...PMID: 25538079. PubMed Central PMCID: PMC4353587. 2. Stephen, Andrew G., et al. "Dragging ras back in ...daily, yet obstacles remain. One of the biggest challenges facing researchers today is the complexity of... of the latest resources? In response to this challenge, Addgene has launched an initiative to curate ...

  9. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    ...Era of Personalized Medicine. Genes Dis. 2017 Jun;4(2):43-63. DOI: 10.1016/j.gendis.2017.04.001 Maetzig...However, the cargo capacity of AAV is ~4.7 kb, almost half that of lenti and gamma-retro viruses. This is large...

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

    Type
    Blog Post
    ... high-priced commercial devices.     Fig. 2: Workflow for open software (left) vs. open hardware...hardware, this process is often much more involved. Challenges such as the provision of an accessible supply...

  11. Hot Plasmids - May 2022

    Type
    Blog Post
    ... those that also infect the liver.    Fig. 2: Summary of the generational evolution of the AAV ...Leeson Multiplexed images in live cells remains a challenge due to multiple factors, including limitations...

  12. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ...3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2. PubMed. Find the plasmids from this publication ...encoded only once, and each <200 bp tRNA-gRNA takes up half the space of a standard pol III-gRNA, which will...

  13. Summer SciComm: Preprints

    Type
    Blog Post
    ... scientific communication ecosystem Figure 2: The scientific ecosystem, enriched by open access...Ettinger CL, Sadanandappa MK, Görgülü K, Coghlan KL, Hallenbeck KK, Puebla I (2022) A guide to preprinting for...

  14. Plasmids 101: Cre-lox

    Type
    Blog Post
    ..., N. and Hamilton, D. 1981. PubMed PMID: 6276558. 2. Site-specific DNA recombination in mammalian cells...in a particular organ, or bypassing embryonic lethality. To facilitate the use of Cre-lox technology,...

  15. Transferable Skills Guide: Creativity

    Type
    Blog Post
    ...to the break room for a sip of water.  Fig 2: Blugene likes to spark creativity in out on the open... needing to brainstorm with a team. One major challenge with these brainstorming sessions is avoiding ... review it. This survey generated 26 ideas, over half of which, when implemented, impacted the entire ...you can, set aside time to prepare for future challenges. This doesn’t have to be time-consuming or even... keeping your science going throughout these challenging times.  Take a Break! If all else fails, one ...

  17. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    ...systems utilize with type I-F Cascades. Figure 2: Cascade complex from CRISPR type I-E. Created with...this complex, developing Cas3 tools has been challenging. It can be difficult to ensure comparable expression..., though consistent expression can still be a challenge. A multitasking Cas Most Cas effectors have one...

  18. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    ...that view (Callahan et al., 2022).     Fig. 2: Students in an MDH CURE replaced select residues ... Biology (Kimball et al., 2020). Benefits and Challenges CUREs inspire students to take ownership of their...experience on their resumes). CUREs are not without challenges. Real research is messy, and it can be difficult...

  19. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...GUS-staining is a very useful visual reporter (Figure 2). The GUS enzyme cleaves a substrate, which will then...transgenic hairy root lines in tissue cultures takes half a year, and generating a transgenic plant can take...Luciferases usually have shorter mRNA and protein half-lives then GUS and are therefore good temporal reporters...

  20. Bioinformatics at Addgene

    Type
    Blog Post
    ...the necessary steps to perform:      Fig. 2: Quick-stepping to a FASTA file   With our outline...300,000 individual nucleotide sequences. More than half of these sequences were generated by Addgene as ...on-site, but now we were presented with a new challenge: automating the bioinformatics steps needed to...

  21. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    ...start selecting and testing systemic capsids! Figure 2 contains a decision tree for selecting a capsid, while... highlighted below the table.     Figure 2: This decision tree can help guide your choice on ...transfer to the adult brain. Nature Biotechnology, 34(2), 204–209. https://doi.org/10.1038/nbt.3440 Goertsen...annurev-neuro-111020-100834 Challis, R.C., Ravindra Kumar, S., Chan, K.Y., Challis, C., Beadle, K., Jang, ...or lumbar spine. Image and caption courtesy of Challis et al., 2022. Where do they come from? Systemic...https://clover.caltech.edu. Figure adapted from Challis et al. 2022. Image created with BioRender.com. ...low doses a regional bias towards the cortex and thalamus emerges. AAV-PHP.eC effectively transduces neurons...

  22. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    ...Antibody Fragments. Antibodies (Basel). 2019 Apr 9;8(2):28. doi: 10.3390/antib8020028. PMID: 31544834; PMCID...reduced-in-size options often have reduced binding strength, half-lives, and/or specificity as well. They’re more ...

  23. RNA Extraction Without A Kit

    Type
    Blog Post
    ...156–159. https://doi.org/10.1016/0003-2697(87)90021-2 Chomczynski P, Sacchi N (2006) The single-step method...until homogenization and cell lysis. This can be challenging depending on your laboratory situation or tissue...

  24. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...of 3,5-difluoro-4-hydroxybenzylidene-imidazolinone-2-oxime (DFHO), a fluorophore modified from DsRed. DFHO...RNA is not inherently fluorescent and can be challenging to track intracellularly, but aptamers permit... when bound to DFHBI despite being approximately half the length (Warner et al., 2014); its length reduces...next additions to aptamer soup! References Alam, Khalid K., et al. "A fluorescent split aptamer for visualizing...

  25. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    ...strain (which will be covered in more detail in part 2 of this blog series) controls the cell types in which...the injection of a targeting vector alone, some challenges still exist. Nucleases may cause off-target effects...

  26. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ...this year. Designing the donor plasmid Figure 2: Donor plasmid design strategy. A donor plasmid contains...Institute website at alleninstitute.org. A classic challenge in cell biology is making sure that what we observe...

  27. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...excise the HIV-1 genome from every infected cell and 2. prevent reinfection of those cells. Kaminski et al...be the way to accomplish this tough job! Kamel Khalili’s lab at Temple University has demonstrated two ... a well-established viral reservoir. A second challenge for CRISPR HIV-1 therapies is sequence specificity..., Kaminski R, Fagan PR, Putatunda R, Young WB, Khalili K, Hu W. CRISPR/gRNA-directed synergistic activation...T, Tedaldi E, Napoli A, Zhang Y, Karn J, Hu W, Khalili K. Elimination of HIV-1 Genomes from Human T-lymphoid...PMID: 26035283. PubMed Central PMCID: PMC4649911. Khalili K, Kaminski R, Gordon J, Cosentino L, Hu W. Genome...

  28. Building Global Connections with the International Mentorship Program USA-EUROPE

    Type
    Blog Post
    ...15.4%) 46 (59%) of these students were in the last 2 years of their degree. Students filled in a pre-program...PhD, achieving our career goals can be a real challenge. On top of that, in the midst of a rapidly changing...mentees with a broader, first-hand vision of the challenges and career opportunities that truly exist outside...was much more than that. I discovered that the challenges that I am facing in my career are common problems... that many others have faced yet, still, this challenge did not hinder them from advancing their careers...

  31. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    .... Find Plasmids from the publication at Addgene 2. Green, Alexander A., et al. "Toehold switches: de-novo-designed...symptoms, including the development of fetal microcephaly and Guillain-Barré syndrome. In response, the...

  32. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...gene1 [lon] ompT gal sulA11 R(mcr-73::miniTn10--TetS)2 [dcm] R(zgb-210::Tn10--TetS) General protein expression...suited for recombinant protein expression. Many challenges can arise when over-expressing a foreign protein...functional RNaseE which results in longer transcript half-life F- ompT lon hsdSB (rB- mB-) gal dcm rne131...

  33. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...improvement in muscle physiological performance.  2. Trans-splicing Overview: For trans-plicing, splice...packaging capacity is limited to ~4.7kb, or roughly half the packaging limits of lentiviral and adenoviral...multiple copies of both vectors. Examples of use: Halbert et al., 2002 used an overlapping vector with 440...heterodimer, then splicing of the pre-mRNA of the two halves of the transgene will reconstitute the full-length...A of the transgene contains the promoter, the 5’ half of the transgene, a splice donor sequence and the... of homology, a splice acceptor sequence, the 3’ half of the transgene. The full length gene is reconstituted...124–130 . https://doi.org/10.1038/sj.mt.6300322 Halbert CL, Allen JM, Miller AD (2002) Efficient mouse ...

  34. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ... PMID: 9537413. PubMed Central PMCID: PMC3010411. 2. Kohli, Manu, et al. "Facile methods for generating...double strand break repair, it has proven more challenging to develop high efficiency CRISPR-based approaches...delivering CRISPR components as RNPs and delivering two halves of a repair template spread across two AAVs, it...

  35. CRISPR Guide

    Type
    Guide
    ...Browse Plasmids: Double-Strand Break (Cut) Figure 2: Overview of the NHEJ repair mechanism. Multiplex ...systems enable researchers to target anywhere from 2–7 genetic loci by cloning multiple gRNAs into a single...Cas9 is included in the gRNA-containing plasmid, or 2-vector systems, in which Cas9 must be delivered separately...your experimental cell population (Figure 8E). In a 2-vector system, you’ll need to either co-infect with...the presence of infectious organisms (like SARS-CoV-2 ) and genetic mutations. Similar to Cas9 and Cas12...Cas enzymes? CasPEDIA is an encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme...off-target effects by using a single Cas9 nickase and 2 different gRNAs, which bind in close proximity on ...

  36. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...You’ve probably heard that only 2% of our genome is made of protein-coding genes, and you might be wondering...in Plants with the CRISPR-Cas Technology is a Challenge yet Worth Accepting. Front Plant Sci. 2015;6. ...

  37. Lentiviral Vector Guide

    Type
    Guide
    ...vectors. Viruses , 3 (2), 132–159. https://doi.org/10.3390/v3020132 PMID: 22049307 Ghaleh, H. E. G., Bolandian... Therapy , 24 (2), 132–142. https://doi.org/10.1089/hum.2012.229 PMID: 23311447 Shalem, O., Sanjana, N...intact. First-generation plasmids include (Figure 2): Transfer plasmid — contains transgene and wild-type...envelope removed Envelope plasmid — contains env Figure 2: First-generation lentiviral plasmids. Created with...plasmid 1 — contains gag and pol Packaging plasmid 2 — contains rev Envelope plasmid — contains env Third-generation...Infectivity and Integration Targeting. PLoS ONE , 2 (12), e1340. https://doi.org/10.1371/journal.pone.0001340...Lentiviral Vector Uses and Overview Overcoming the Challenges of Lentiviral Production Your Lentiviral Plasmid...

  38. Of Myc and Men

    Type
    Blog Post
    ...promising monoclonal antibody with high specificity. (2) Narrow down the specific epitope that your antibody...in these species, they found that the N-terminal half of the antigen differs significantly between human...

  40. Adenovirus Guide

    Type
    Guide
    ...use of pAdEasy-2 (Addgene #16401) can increase the capacity of the rAdV vector. pAdEasy-2 does not contain... incorporate additional genomic deletions. Figure 2: First-generation rAdV vectors. Created with BioRender.com...and have to be handled at biosafety level two (BSL-2). They are highly immunogenic and can trigger a strong...era of personalized medicine . Genes & Diseases, 4 (2), 43–63. https://doi.org/10.1016/j.gendis.2017.04.001...adenoviruses using the AdEasy system . Nature Protocols, 2 (5), 1236–1247. https://doi.org/10.1038/nprot.2007.135...Adenoviral vector vaccine platforms in the SARS-CoV-2 pandemic . NPJ Vaccines, 6 (1), 97. https://doi.org...Adenoviral backbone plasmid that lacks E1 and E3. pAdEasy-2 Adenoviral backbone plasmid that lacks E1, E3, and...
Showing: 241 - 280 of 303 results