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Showing: 81 - 100 of 207 results
  1. Viral Vectors 101: Voltage Indicators

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    Blog Post
    ...depths (1 mm), important for certain applications such as in vivo tissue imaging.   Figure 1: Ace-mScarlet...sources – voltage-gated potassium channels found in Drosophila to phosphatases from sea squirts. This design... detector in this system was dipicrylamine, a lipophilic compound that distributes itself within the plasma...sensing) hydrolyzes a precursor dye to make it more lipophilic, increasing its affinity for the membrane, where... fluorescent component (i.e., when the dye is lipophilic and integrates into the membrane), the voltage...
  2. Plasmids 101: Knockout/Knock-In Plasmids

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    Blog Post
    ... the example shown in Figure 1, large sequences corresponding to exons 1 and 3 of the targeted gene have...knock out exon 2 of a given gene.   Figure 1: A knockout targeting vector designed to insert a ... the Neomycin resistance (NeoR) gene between exon 1 and 3 of the targeted gene. Upon recombination, the...can be used to replace genes in yeast, mice, or Drosophila through homologous recombination. The concept...
  3. Celebrating One Million Plasmids Shared!

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    Blog Post
    ... @Addgene congrats for 1 million orders. @RothLabUNC accounts for 10,000 or 1%! — BryanRoth (@zenbrainest... scientific sharing community (check out the infographic below for stats on our plasmid deposits and requests...
  4. Cpf1 Update: Comparison to Cas9 and NgAgo

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    Blog Post
    ...on-target and off-target editing efficiency Figure 1: Effects of (A) double and (B) single base pair gRNA-target...that Cpf1 can tolerate mismatches at gRNA positions 1, 8, 9, and 19-23. Accordingly, the 3’ end of the gRNA...Most gRNAs directed low-frequency Cpf1 cleavage at 1-12 off-target sites; in contrast, SpCas9 may cleave...fits into the genome editing landscape. References 1. Kim D, Kim J, Hur JK, Been KW, Yoon SH, Kim JS. Genome-wide... off-target editing rates, determined using sophisticated genome-wide analysis. At many of its computationally...
  5. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

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    Blog Post
    ...attract fish for them to prey upon (Figure 1). Figure 1: The sea anemone's red color can be used ...synthetic biology. Journal of Biological Engineering, 12(1). https://doi.org/10.1186/s13036-018-0100-0 Murakoshi...expression in living cells. Scientific Reports, 9(1). https://doi.org/10.1038/s41598-019-48604-4 Salih...surface of the chromoprotein Ultramarine with hydrophilic amino acids to reduce non-specific interactions...
  6. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

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    Blog Post
    ...discovered such as Kaede and PS-CFP2 (see Table 1). Table 1: Properties of Selected Irreversible Photoactivatable...) by mutating Threonine 203 to Histidine (T203H) (1). wtGFP normally contains a mixed population of neutral...in fluorescence when illuminated with blue light (1). Similar to PA-GFP, scientists later developed red...fixation conditions to preserve cellular structures (0.5-1% osmium tetroxide). These conditions destroy most ...with extraordinary molecular detail.   References 1.Patterson, George H., and Jennifer Lippincott-Schwartz...is believing." Annu. Rev. Biomed. Eng. 10 (2008): 1-38. PubMed PMID:18647110. Additional Resources on ...reversible photoactivation - and have enabled sophisticated imaging techniques.     Irreversible photoactivation...
  7. COVID-19 Resources

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    Collection
    ...mediating the entry of SARS-CoV and SARS-CoV-2 viruses. ( 1 ) TMPRSS2 - a serine protease that primes the SARS-CoV...transcriptase expression plasmids from the Drew Endy and Philippa Marrack Labs. Point-of-care testing for COVID-...
  8. Plasmids 101: Using Transposons in the Lab

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    Blog Post
    ...Transposons: The jumping genes. Nature Education 1(1):204. https://www.nature.com/scitable/topicpage/transposons-the-jumping-genes... They transpose by a ‘copy-paste’ mechanism (Fig. 1). They first copy themselves as RNA transcripts and...their host’s genome.   There are two types of Class 1 TEs: those with long terminal repeats (LTRs) and those...reverse transcriptase activities.    Figure 1: Overview of retrotransposon transposition. Retrotransposons...CRISPR-Cas system. This system has four major components: 1) a CRISPR RNA (crRNA), 2) four proteins (TniQ, Cas8...lentivirus). Large cargos, such as the ~11 kB cDNA for dystrophin, the gene mutated in muscular dystrophy, require...
  9. Hot Plasmids: Winter 2025

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    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...Park, J. C., Zhao, J., Luebbers, A., Green, E., Philibert, C. E., Zhang, H., Layne, M. D., Wu, J. C., & ...
  10. Five Popular Model Organisms, Part 2

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    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..., or frogs, are one of the most popular model amphibians in science with early research providing insight... Jiang, J. P. (2016). Research proceedings on amphibian model organisms. Dong wu xue yan jiu = Zoological...
  11. FlipGFP, a novel fluorescence protease reporter to study apoptosis

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    Blog Post
    ...central ɑ-helix that can be split into 3 parts: (1) β-strands 1-9 (β1-9) and the one ɑ-helix, (2) β-strand ...apoptosis in HeLa cells, zebrafish embryos, and Drosophila. They treated HeLa cells expressing FlipGFP with...visualize apoptosis of enterocytes in the midgut of Drosophila, an observation that has been difficult until...
  12. New Tools Enable CRISPRa for Neuroscience Applications

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    Blog Post
    ...by mating with Cre-expressing mice. Figure 1: (A) Schematic of the Cre-dependent SPH construct ... alpha (aCaMKII; excitatory neurons) and Synapsin-1 (pan-neuronal) promoters (Figure 1b), and sgRNAs can...ubiquitous expression of sgRNA(s) and mCherry (vector 1) with neuron-specific expression of dCas9-VPR (Figure...Neuron-specific expression is mediated by the synapsin 1 (SYN) promoter, which was superior in its ability ...immediate-early gene Bdnf, which encodes brain-derived neurotrophic factor (BDNF). Bdnf is expressed early in response...
  13. A Needle in a Base-Stack: Cas9 Structural Biology

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    ... basic parts of the Cas9 enzyme.   Figure 1: A cartoon depiction of Cas9’s two major lobes, REC...the CRISPR system (Nishimasu et al., 2014). Table 1. Summary of Cas9 domains, the lobe each domain belongs...state (PDB ID 4CMP) with domains colored as in Figure 1. HNH, red; RuvC, orange; CTD, yellow; REC lobe, light... on itself at the bottom of the image. Stem loops 1 and 2 extend horizontally across both lobes. The guide...outside of the protein, spanning both lobes. Stem loop 1, nestled between the REC and CTD domains, is required...Within an interior cavity, Cas9 grips nucleotides 1-10 of the guide RNA, prearranged for base pairing ...343–347. https://doi.org/10.1038/s41586-022-04470-1 Cofsky, J. C., Soczek, K. M., Knott, G. J., Nogales...
  14. Viral Vectors 101: Viruses as Biological Tools

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    ...unleashes its cargo – its own genome.   Fig. 1: Viral particle infection into a host cell. Image ...Vector Resources at Addgene! References and Resources 1. Roe T, et al. “Integration of murine leukemia virus...nuclear import of human immunodeficiency virus type 1 preintegration complexes.” Proc. Natl. Acad. Sci. ...approved in the US for multiple diseases, including hemophilia B and spinal muscular atrophy.   Type of Virus...
  15. Supporting Reproducibility with a Connected ELN

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    ...true reproducibility. The following are critical: (1) connectivity to other data sources used in research...engagement with the Wisconsin research community were to: (1) prevent vendor lock in by making it easy to get data...other large research institutions. The following graphic depicts the ways in which the two guiding principles...it difficult or impossible to reproduce. As the graphic depicts, RSpace supports linking to all of the ... other tools and resources. As depicted in the graphic, RSpace is integrated with most of the tools noted...
  16. The PAM Requirement and Expanding CRISPR Beyond SpCas9

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    ... modify any desired genomic target provided that (1) the sequence is unique compared to the rest of the...will cover three ways to circumvent this limitation: 1) the use of novel S. pyogenes Cas9 variants with varying...Cas9 from Staphylococcus aureus (SaCas9) is about 1 kilobase smaller than SpCas9, so it can be packaged...Biol 19: . https://doi.org/10.1186/s13059-018-1591-1 Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, ...Campylobacter jejuni (Cj) NNNNRYAC Streptococcus thermophilus (St) NNAGAAW Treponema denticola (Td) ...
  17. Deep Dive: Statistical Tests (Comparisons)

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    Blog Post
    ... closely data points fall to each other (see Fig. 1). Alpha values – numerical risk allowance in a ...statistical test assumes your data follows. Fig. 1: Examples of comparable groups with and without equal...values from smallest to largest and ranking them (1 being the smallest value). It calculates the cut off...directly manipulating affect an outcome (age, demographic, diet, etc., in a cancer drug clinical trial,...know type of distribution you are working with, graphing data with a histogram is sometimes sufficient ...
  18. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

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    Blog Post
    ..., including those found at Addgene! Figure 1: Strategy for assembling the barcoded gRNA library...pool-clone them into the pAWp28 storage vector (Figure 1): Forward oligo: 5’- CACCGNNNNNNNNNNNNNNNNNNNGTTTGGGTCTTCGAGAAGACCTATTCXXXXXXXXC... He can be contacted at [email protected].   References 1. Cheng, Allen A., Huiming Ding, and Timothy K. Lu....26190106. PubMed Central PMCID: PMC4559080. 8. Shalem, Ophir, et al. "Genome-scale CRISPR-Cas9 knockout screening...
  19. Plasmids for Endogenous Gene Tagging in Human Cells

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    Blog Post
    ...Collection Fluorescent tagging strategy Figure 1: Gene editing is the addition of a useful coding sequence...plasmid design strategy. A donor plasmid contains 1 kb homology arms matching the gene of interest, a ...Institute website at alleninstitute.org. References 1. Roberts, Brock, et al. "Systematic gene tagging using...includes systems for tagging mammalian cell lines, Drosophila cells, and C. elegans. In addition to generating...
Showing: 81 - 100 of 207 results