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We narrowed to 603 results for: RAN-1

Showing: 181 - 200 of 603 results

  1. The Materials Science of Optogenetics Experiments

    Type
    Blog Post
    Published
    Sept. 17, 2015, 2:30 p.m.
    ... 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 ...tissue, placement above the site of stimulation guarantees that the majority of the opsins will receive ...tangled. Furthermore, to give the animal the full range of motion, a commutator allows the patch cable to...

  2. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    Published
    July 14, 2016, 2:30 p.m.
    ...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...pattern opens up the possibility of directional gene transfer, analogous to traditional restriction enzyme cloning... cloning. Sticky-end mediated gene transfer would be particularly helpful for targeting non-dividing cells...gene delivery due to their low immunogenicity and range of serotypes allowing preferential infection of ...

  3. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    Published
    Jan. 11, 2017, 8:47 p.m.
    ...Protocols Publication describing them here. References 1.Ran, F. Ann, et al. "Genome engineering using the CRISPR-Cas9...plasmid post transfection. The 2A "self cleaving" peptide causes the ribosome to translate Cas9-2A and ...component of most CRISPR systems and generates double strange breaks at target sites 5' of the 5'NGG3' PAM sequence...backbone. This particular backbone can be used to transfect a variety of mammalian cells and can be modified...are some additional features. The first and most imporant is that, upstream of the Cbh promoter, this plasmid...can be used to test for SpCas9 expression post transfection by western blot. Overall, this is a versatile...

  4. Transferable Skills: Negotiation

    Type
    Blog Post
    Published
    March 3, 2022, 2 p.m.
    ...dollar contracts behind closed doors. Fig. 1: The mythical land of negotiations.    You negotiate...feel they lack formal training in a variety of transferable skills. At Addgene, we've set out to fill this... by highlighting that researchers learn many transferable skills while working in the lab and by offering...where you might need some help. Today in our transferable skills guide: Negotiation. As a grad student...I was negotiating with my PI on how to spend my grant money (Can I attend this conference?) and with my...Hawaii?). So how can you approach improving this transferable skill? Negotiation as a zero sum game Some ...is the 6th year grad student frustrated that the brand new grad student keeps using the machine? Let...

  5. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ...genome replication and transcription of negative strand RNA viruses.   References 1. Buchholz, Ursula J....which are required for replication and transcription (Figure 1). All viral genes are removed and replaced...negative strand RNA viruses: The power of reverse genetics. Springer Berlin Heidelberg, 2004. 1-41. PubMed...cells that stably express T7 (e.g. BSR T7/5 cells) (1). We hope that minigenomes will greatly enhance our...gene transcription (6). The EBOV genome contains seven viral genes, each flanked by a transcription start...replicated and transcribed in the cell when the appropriate viral proteins are provided in trans (see Other...genes and the minigenome are transcribed (and the viral mRNAs are translated) by cellular machinery. NP ...

  6. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ...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 ...engineered combinations of transcriptional activators to induce gene transcription at genomic sites targeted...p65-HSF1 transcriptional activator to each genome-bound dCas9 (see Figure 1a for the transgenic construct...different untranslated or coding regions. Typically, each gene isoform has a distinct transcriptional start...

  7. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ...cells need three different plasmids to produce AAV: 1) the RepCap plasmid which provides the AAV replication...plasmid. This whole process takes ~3 hours to complete: 1 hours hands-on time and 2 hours for the qPCR run and...about AAV production and purification! References 1. Aurnhammer, C., Haase, M., Muether, N., Hausl, M.... repeat sequences. Human gene therapy methods, 23 1, 18-28. PubMed PMID: 22428977. 2. Choi, V.W., Asokan...proteins to process a double-stranded intermediate into the single-stranded genome that's then packaged...viral replication; and 3) the transfer plasmid which encodes a transgene of interest that’s packaged into...XmaI restriction sites). Another way is to transform AAV transfer plasmids into bacterial strains, like NEB...

  8. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    Published
    April 26, 2022, 1:15 p.m.
    ...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...pipetting error or an incredible discovery. The range of RQ values used to call copy numbers should be...fluorescent dye that works by binding to all double-stranded DNA, while Taqman uses an intercalating dye, with...confident determination of copy number, albeit within a range of RQ values as noted above.   In both applications...

  9. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    Published
    April 25, 2017, 2:30 p.m.
    ...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 ...fluorescent PA-FPs by performing several rounds of random mutagenesis screening for enhanced RFP variants...

  10. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ... emailed submission) in the Deposit menu: Figure 1: Begin the deposit process by clicking "Submit Online...the plasmids, or (for deposits submitted after July 1, 2025) after three years on hold, Addgene staff may...cases Please contact us at [email protected] or +1 (617)-ADDGENE (233-4363) if your deposit falls into...Addgene?" or contact us at [email protected] or +1 (617)-ADDGENE (233-4363) with additional questions...strain. Please contact us at [email protected] or +1 (617)-ADDGENE (233-4363) if you’d like to distribute...page so that any data you enter is not lost.   Step 1: Sequences, Maps, and Files Please provide any full...a 1.5 mL microfuge tube at a concentration of 0.1–1 µg/µL (or bacterial streaks, if you prefer). If you...

  11. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Published
    July 31, 2018, 12:55 p.m.
    ...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...and they had been shown to to transduce the retina but only transiently. However new viral vectors capable...adenovirus (AdV) and lentivirus (LV) are known to transduce retinal cells, AAV quickly became the vector of...due to its considerable immunogenicity and its transient expression. LV results in long term expression... in dividing and non-dividing cells due to its (random) integration into the genome, and its large DNA...

  12. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    Published
    Feb. 4, 2025, 2:15 p.m.
    ..., 10(1). https://doi.org/10.1038/s41467-019-12449-2 Pupo, A., Fernández, A., Low, S. H., François, A.,...therapy of human diseases. Signal Transduction and Targeted Therapy, 9(1). https://doi.org/10.1038/s41392...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...vivo starts with transducing cells with the CRISPR library in vitro before transplanting them into mice....time. This reduces the chance of integration at random locations that could cause deleterious effects....portions of the AAV genome can integrate at double-stranded break points caused by CRISPR-Cas9 editing. The...

  13. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    Published
    May 30, 2019, 12:44 p.m.
    ...identification of proteins associated with the IRF-1 locus. Figure 1: Schematic of enCHIP. Image from Fujita... interest, the promoter region of the transcription factor IRF-1. Immunoprecipitation, using an antibody...References Campbell, Amy E., et al. "NuRD and CAF-1-mediated silencing of the D4Z4 array is modulated ...dCas9 has focused on transcriptional activation using a fusion to a transcriptional activator such as VP64...repression of transcription through binding a promoter region to inhibit association of transcriptional activators...2018). Furthermore, the Fujii Lab generated a transgenic mouse line and retroviral expression system for...Tapscott Lab identified regulators of DUX4, a transcriptional regulator that when inappropriately expressed...

  14. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ...10.1038/s41551-024-01227-1. doi:  https://doi.org/10.1038/s41551-024-01227-1   CHARM: A compact epigenetic...spacer proteins ending with a GFP nanobody (Figure 1). This enrichment streamlines the purification and...highly-visible beads on the cryo-EM grid.     Figure 1: MagIC-cryo-EM for sample enrichment and structure...single-particle analysis of PRC2 with RvLEAMshort (1:6 molar ratio) with 10 minutes of glutaraldehyde crosslinking...which ensures the editor will only be transiently present in transduced cells, limiting off-target editing... dCas9 fused to a DNA methyltransferase — that can drive lasting transcriptional silencing (Nuñez et al...virus-like particles (eVLPs) offers an option to transduce cells directly with ribonucleoprotein cargo. eVLPs...

  15. Expanded Access to Rabies Plasmids

    Type
    Blog Post
    Published
    Feb. 1, 2024, 2:15 p.m.
    ... export license applications for VSV and SARS-COV-1 plasmids. Export control license process    An export...charged.  Timelines  We have offered VSV and SARS-COV-1 plasmids under this application process since May ...where Addgene is located.  The export license, if granted, allows us to ship rabies plasmids and viral vectors...

  16. Hot Plasmids: Winter 2025

    Type
    Blog Post
    Published
    March 11, 2025, 1:15 p.m.
    ...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...NC: nucleocapsid RNA chaperone, RT: reverse transcriptase. Right: T1 tomato plants that inherited an edited...Cultured rat hippocampal neuron expressing pan-membrane ASAP5. Solid blue circle marks the initiation ...via the Kv2.1 targeting motif as well as a pan-membrane construct without the Kv motif. Altogether, if...

  17. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    Published
    Aug. 3, 2023, 1:15 p.m.
    ...markers in the panel. This complexity index ranges from 0-1, with a value of “0” indicating those markers...detector for that fluorophore. For example, in figure 1, the FITC signal is spilling over into the primary...spectra to use for compensation.     Figure 1: The emission spectra and filters for FITC and PE ...completely distinct from one another, and a value of “1” indicating those markers are identical and completely...

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

    Type
    Blog Post
    Published
    May 24, 2016, 2:30 p.m.
    ... here! Podcast Breakdown (Time: Topic) 0:00-1:00 : Intro 1:01-3:48 : Kingdom Kwapata's background and ...Michigan State University 9:36-10:29 : Kingdom's transition back to Malawi and Lilongwe University of Agriculture...-14:53 : Kingdom's current projects producing transgenic crops 14:54-23:44 : Follow up Kingdom's PhD projects... transgenic crops...

  19. Our New Antibody Mascot is....

    Type
    Blog Post
    Published
    Feb. 24, 2022, 2:15 p.m.
    ...introduce their new friend…  Fig. 1: Meet...well, frankly, we're not sure yet!   This as-yet–...narrowed it down to three choices.   Fig. 1: Lin, Abi, or Blottie? Which one suits me best?   ...choices), we'll be in touch with you soon with some brand-new Addgene swag!   Last month, we introduced you...out to you and received a plethora of options, ranging from Affie to Yncredibody. After an agonizing week...

  20. Multiplexed Overexpression of Regulatory Factors (MORF) collection

    Type
    Blog Post
    Published
    March 30, 2023, 1:15 p.m.
    ... Fig. 1: Barcoded TF library was packaged in lentiviral particles and used to transduce human embryonic... J. et al., A transcription factor atlas of directed differentiation. Cell. 2023. 186 (1) 209-229. doi...TF library, with each cell receiving only 1 TF expression construct. Cells were cultured following a differentiation...Transcription factors (TFs) act as the gate keepers for gene expression, turning transcription on and... regulatory network. The transcription atlas and library The transcription factor library The human...approximately 1,800 TFs, with >3,500 unique isoforms transcribed from these sites. The Zhang lab generated a comprehensive... contribute to differentiation, the Zhang lab transduced human embryonic stem cells with the viral TF ...
Showing: 181 - 200 of 603 results