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We narrowed to 328 results for: hal.1

Showing: 121 - 140 of 328 results

  1. Optogenetics Guide

    Type
    Guide
    ...algae Chlamydomonas reinhardtii . Channelrhodopsin-1 (ChR1) is excited by blue light and permits nonspecific...based on the LOV2 domain of Avena sativa phototropin 1 LOVETRAP reversibly sequester and release proteins...concept of optogenetics. 2012 Prog Brain Res. 196: 1-28. PMID 22341318 Gradinaru V, Zhang F, Ramakrishnan...diversifying and extending optogenetics. Cell. 196:1-28. PMID 20303157 Han X, Boyden ES. 2007 Multiple-...Limitations and Future Developments. Exp Physiol. 96(1): 19–25. PMID 20621963 Mattis J, Tye KM, Ferenczi ...plasmids . Halorhodopsins Halorhodopsins are light-gated inward chloride pumps isolated from halobacteria. Wild-type...Jaws Browse Halorhodopsin plasmids . Archaerhodopsins Archaerhodopsin-3 (Arch) from Halorubrum sodomense...

  2. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    ...events by HDR can be generated more efficiently by (1) improving editing accuracy by preventing re-editing...specifically incorporate homo- or heterozygous mutations. 1. Increasing HDR accuracy with CRISPR/Cas-blocking ...targeted alleles when gene editing.   Figure 1: Techniques to improve editing efficiency.   But...provided by blocking mutations. Essentially, you:  Step 1 - Introduce the blocking mutation together with the...developing novel iPSC-based model systems.     References 1. Cong, Le, et al. "Multiplex genome engineering using...distance is only 10 bp, the efficiency already drops by half and after only about 30 bp, it is no longer feasible...

  3. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    ...following four components into 293T cells. Figure 1: The key components of Nanoblades. Image from Mangeot...protein. A single Nanoblade particle can be loaded with 1-4 different gRNAs. Viral envelope proteins To alter... between ~50-70% for these four cell types (Table 1). Additionally, hiPSCs and mouse BM cells still behaved...hepatocytes  Myd88  ~50% Human HSCs  Myd88  ~50% Table 1: Nanoblades editing efficiencies of primary cells....pronucleus of the zygote which is both technically challenging and can negatively impact zygote viability. Nanoblades...

  4. The Materials Science of Optogenetics Experiments

    Type
    Blog Post
    ... different steps and materials required for each [1, 2]. This protocol [3] provides, in exquisite detail...follow him on Twitter @derekpsimonphd.    References 1. Gutierrez DV, et al. Optogenetic control of motor...manipulation of neural circuits. Nature protocols. 2012;7(1):12-23. doi: 10.1038/nprot.2011.413. PubMed PMID: ...freely moving mice. Neuroscience research. 2011;70(1):124-7. doi: 10.1016/j.neures.2011.01.007. PubMed ...validation experiments we discussed last week are only half of the battle when using optogentics to answer a...

  5. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...oxygen. Additionally, its status as a Biosafety Level 1 microbe makes it a perfect candidate for introductory...Transport, biofilms, and much more. Because of it’s BSL-1 status, E. coli can be used as a model to relate these...with superior performance. Nature Communications, 9(1). https://doi.org/10.1038/s41467-018-04899-x Yokoyama...biomedical model organism.  Arabidopsis thaliana Arabidopsis thaliana is one of, if not the most, recognized...Arabidopsis genetics research development. Arabidopsis thaliana was fully sequenced by the year 2000, becoming...properties affecting adaptations.  Arabidopsis thaliana flowers. Image from BlueRidgeKitties. If you...functions in an ecological context with Arabidopsis thaliana. ELife, 4. https://doi.org/10.7554/elife.06100...

  7. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ...some technical obstacles when working with virus: 1. Viral vectors undergo DNA rearrangements. Depending...approximately 20–50% of the packaging cells after 1–3 days of selection. A lower percentage of surviving...up-to-date tips for your virus experiments. References 1. Anthony, Simon J., et al. "A strategy to estimate...tasks, both of which are factors in the technical challenges they can present. Many researchers find themselves...after transfecting cells, replace the medium with half the volume typically used (e.g., use 5 mL medium...

  8. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ... spread out onto the plate. We test the (capped) 1:1 agarose/DMEM solution against a part of the body ...a 60 mm dish ready for picking is shown in figure 1.   Find Additional Useful Protocols on Addgene's Protocol...incubator, additional growth medium is added to about halfway up each well. Tips for using the agarose cloning...Time CRISPR User (by a 1st Time CRISPR User) The Challenges of Cell Culture Plasmids 101: Mammalian Vectors...

  9. Podcast: A Malawian Professor's Path to Biotech Research

    Type
    Blog Post
    ... here! Podcast Breakdown (Time: Topic) 0:00-1:00 : Intro 1:01-3:48 : Kingdom Kwapata's background and ...Kingdom's PhD projects - HIV inhibitors, biofuels, challenges in Malawi, and more 23:45-28:07 : How Seeding...

  10. CRISPR Guide

    Type
    Guide
    ... most popular genome engineering approach. Figure 1: Overview of the basic CRISPR mechanism Engineered...from Staphylococcus aureus ) has a coding sequence ~1 kb shorter than SpCas9 while retaining the same basic...lentiviral transfer vectors (Figure 8B). Libraries come in 1-vector systems, in which Cas9 is included in the gRNA-containing...efficiency and fidelity. Nature Communications , 13 (1), 1425. PMID: 35301321 Edraki, A., Mir, A., Ibraheim...interrogation by SpRY-Cas9. Nature Communications , 15 (1), 3663. PMID: 38688943 Hsu, P. D., Scott, D. A., Weinstein...increase its specificity. Nature Communications , 9 (1). PMID: 30082838 Maddalo, D., Manchado, E., Concepcion...CRISPR-Cas9 with Bacteriophage Proteins. Cell , 168 (1–2), 150-158.e10. PMID: 28041849 Sakuma, T., Nishikawa...

  11. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...dsRNA via an RNase III enzyme named Dicer (Figure 1). The plant kingdom has expanded its repertoire of...in the biogenesis of different small RNAs (Figure 1). These siRNAs can exert their gene silencing effect...modifications of complementary gene sequences (Figure 1). As a result, gene silencing can be easily induced...the ease of use in dsRNA applications. Figure 1: Schematic of an RNAi-inducing transgene. Typically...complementary to your gene target as inverted repeats (Figure 1). With this technique, multiple genes can be simultaneously...selection of plants undergoing RNAi, and LIIbeta F 1-2 RNAi, which allows assembly of intron-spliced hpRNAs...such as in the model flowering plant Arabidopsis thaliana (Li et al., 2013; Zhang et al., 2018) and the ...

  12. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ...against the limitations of the assay.      Fig. 1: A schematic representation of Sybr Green chemistry...Methods Mol Biol. 2018;1768:143-160. doi: 10.1007/978-1-4939-7778-9_9. PMID: 29717442. Arya M, Shergill IS...amplification of Plasmodium falciparum multidrug resistance-1 gene (pfmdr1). J Parasitol. 2011 Oct;97(5):939-42....gamma receptor genes FCGR3A and FCGR3B. PloS one, 10(1), e0116791. https://doi.org/10.1371/journal.pone.0116791...available, ddPCR may be a better option for more challenging cases.  Price point If budget is a concern...

  13. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    ...2021).   Table 1: Properties of modern NIR FPs     NIR FP Ex, nm Em, nm ECa, M-1cm-1 QYb, %   Molecular...expression level. State-of-the-art FPs (listed in Table 1) were optimized for efficient binding of endogenous...previously reported widely used dimeric iRFPs (Table 1). In general, the monomeric state of a probe leads...iRFPs in cellular and molecular brightness (Table 1).  NIR reporters and biosensors NIR reporters can ...NIR. Usually, mercury lamps are suboptimal, metal halide lamps are better, and xenon lamps are the best ...

  14. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ... to other technologies like CRISPR. Figure 1: An overview of the general schematic of CRISPR/Cas... can be detected – in the zeptomolar range Time: ~1 hour Equipment required: 37°C water bath and 42°C ...nearly single-molecule sensitive. In the case of HIV-1 and SARS-CoV-2 detection, AIOD-CRISPR without preamplification... Not mentioned in the paper, likely approximately 1-2 hours Equipment needed: Mini PCR machines by DIY-Bio...Sensitivity: – 110 femtomolar  Time: Not mentioned, likely ~1-2 hours Equipment needed: PCR machines/Heat blocks... This test can be performed in two steps for challenging samples as in Two-Step-SHERLOCK or in a fast,...

  15. 5 Steps to Writing Grant Proposals for Grad Students

    Type
    Blog Post
    ...offers insight into a career as a Professor. Step 1: Your PI forwards you a Request For Applications (...proposal typically lists the deliverables, explains 1-2 potential pitfalls of the intended protocol, and...while writing each section. Research challenge The Research Challenge motivates the request for application...how you, the expert, will address the research challenge. Now is the time to make bold statements. While...solution is the best approach to the research challenge. Demonstrating expertise in the field and past...how your innovation will address a variety of challenges and impact the field - its encouraged to dream...time budget writer, you may find this section challenging because it’s hard to determine the exact value...

  16. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    ...determined LY6A, also known as stem cell antigen-1 (SCA-1), is the receptor used by PHP.B to reach the CNS...William L. Stanford. "Concise review: stem cell antigen‐1: expression, function, and enigma." Stem cells 25.6...permissiveness. The first generation of outbred mice (F1) had half the level of CNS PHP.B transduction as their C57BL...check out these two references: Dayton, Robert D., Mychal S. Grames, and Ronald L. Klein. "More expansive...

  17. Antibodies 101: Avoiding the Mouse on Mouse Mess in IHC

    Type
    Blog Post
    ...following three strategies might help!     Figure 1: Species on species staining can be a problem because...same host species as your secondary antibody since 1) your secondary should not recognize Fab fragments...staining can be a challenge in the best of cases. But it can be particularly challenging when your only ...

  18. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ...new combinations of PE protein components (Figure 1), an altered epegRNA delivered with an enhanced pU6cm...amplification of cargo, and heat treatment.  Figure 1: Left: recombined prime editing tools PE2max-NC and...conformational dynamics of the motor protein kinesin-1, the Twelvetrees Lab took advantage of the self-labeling..., Craggs, T.D., & Twelvetrees A.E. (2024) Kinesin-1 is highly flexible and adopts an open conformation... developed in human cells, has been uniquely challenging to apply in dicotyledonous plants, or dicots....

  19. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...NGS) to identify genes of interest. Figure 1: Overview of in vitro AAV CRISPR screens. Created ...certain experiments. AAV can be safely handled at BSL-1, while lentivirus requires a BSL-2 setup. This requires...CRISPR-induced DNA breaks. Nature Communications, 10(1). https://doi.org/10.1038/s41467-019-12449-2 Pupo,...diseases. Signal Transduction and Targeted Therapy, 9(1). https://doi.org/10.1038/s41392-024-01780-w Additional.... V., Teyssier, N., Pan, A., Danner-Bocks, S., Sanghal, T., Yadanar, L., Tian, R., Ma, K., Palop, J. J...

  20. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...and Therapeutic Purposes. Annu Rev Virol. 2014 Nov;1(1):427-51. DOI: 10.1146/annurev-virology-031413-085355...personalized medicine. References and further reading 1. FDA approves gene therapy for blindness 2. Gene Therapy... Med Biol. 2014; 801: 687–693.  doi:  10.1007/978-1-4614-3209-8_86. PubMed PMID: 24664759. PubMed Central... completing her PhD and medical training in Ophthalmology. Gene therapy was in its infancy, but the eye...
Showing: 121 - 140 of 328 results