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We narrowed to 867 results for: FER

Showing: 241 - 260 of 867 results

  1. Plasmids 101: Biotinylation

    Type
    Blog Post
    Published
    Nov. 15, 2018, 1:50 p.m.
    ...solubility, and thus different reagents are useful for biotinylation in different microenvironments. There...used today in molecular biology for an array of different techniques and protocols. In this post we will...and decarboxylases. These enzymes catalyze the transfer of carboxyl groups to organic acids to produce...relatively small, water soluble molecule that does not interfere with the macromolecules it is added to. Biotin... signal and easier detection. This strategy is referred to as the Avidin-Biotin Complex (ABC) immunohistochemistry...variety of biotinylation reagents available that differ in the functional groups or residues they target...a solid substrate (Holden and Cremer, 2003). References 1. Chapman-Smith, A., & Cronan, J. E. “Molecular...

  2. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ..., protected samples and was compatible with the buffer additive MgCl2 and the crosslinking agent glutaraldehyde...PEs) via engineered virus-like particles (eVLPs) offers an option to transduce cells directly with ribonucleoprotein...editor consisting of dCas9 fused to a DNA methyltransferase — that can drive lasting transcriptional silencing...Histone tail for Autoinhibition Release of Methyltransferase (CHARM), with several advantages over the ...  Rather than overexpressing an exogenous methyltransferase for epigenetic silencing, which can be toxic...tail to recruit and stimulate the endogenous methyltransferase. The strategy is generalizable to a variety...D3L recruits the endogenous, autoinhibited methyltransferase DNMT3A to the genomic location specified by...

  3. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    Published
    May 13, 2025, 1:15 p.m.
    ...by Addgenie Rebecca Hutcheson.   References and resources References Banasik, M. B., & McCray, P. B., ...second-generation system as an example.  The lentiviral transfer plasmid encodes your insert of interest, flanked...then, numerous variants have been developed with different mechanisms to inactivate the integrase.  The IDLV...vectors is random, and the integration site will be different for each transduced cell, there’s no telling which... that of regular lentiviral vectors. The only difference is that you would use the integrase-deficient...that titering these IDLVs may require a slightly different protocol than regular lentiviral vectors. This...quantifying the integrated viral DNA along with a reference cellular gene. When titering IDLVs, there is no...

  4. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    Published
    Dec. 6, 2022, 2:15 p.m.
    ...sensors with different ITAM/SH2 reporter pairs allow multiplexed measurement of different RTKs. Although...response. They generated variants optimized for different applications (v82 has higher affinity for glutamate...Measuring local temperature variations and heat transfer at the subcellular level requires a probe with...By fusing two bright fluorescent proteins with different thermal quenching rates (mNeonGreen and tdTomato...sub-millisecond resolution and sub-degree temperature differences under a variety of imaging conditions (155 μs...fast measurement of temperature and local heat transfer. Image reproduced from Lu et al. 2022, under CC-BY-NC-ND...plasmids here!   Lu K, et al. Intracellular Heat Transfer and Thermal Property Revealed by Kilohertz Temperature...

  5. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    Published
    May 20, 2021, 1:15 p.m.
    ...Addgene, I was about to have a check-in with my boss Jennifer. I had finally found my footing with the company... for what we thought would be a couple months. Jennifer and I spent about an hour that morning scrambling... involved multiple people in tight locations. Conference rooms for on-screen presenters! Labs and benches...know cameras, but we needed someone else. Design: Jennifer thankfully volunteered, and we decided to focus...action would be to use an iPad to shoot.  Build: Jennifer created a mobile camera area that included iPad...Zoom, everyone’s new favorite third-party video conferencing software. Interviewees could simply boot up ...while the camera is here, the operator is in a different room.   Design: The previous cycle made me...

  6. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    Published
    March 1, 2016, 3:30 p.m.
    ...scientists have developed and fine tuned many different ways to clone DNA fragments which have provided...more and more common, and for good reason. They offer many advantages over the traditional restriction...with an enzyme master mix, which contains three different enzymes: an exonuclease, which chews back the... this doesn’t allow for mix and match of many different fragments. One way around this is to use a combination...kit you will be using. Some of these products may offer advantages that Gibson assembly does not such as...with a G!) but actually work by significantly different methods. Golden Gate cloning does result in the...from one vector to another. There are lots of different ways to clone these days. Gibson and the other...

  7. Fostering communication in remote collaborations

    Type
    Blog Post
    Published
    April 21, 2020, 1:14 p.m.
    ...are communicating across disciplines or doing different tasks. But these challenges can be exaggerated...equal opportunity to contribute. Using video conferencing tools like Zoom or Google Hangouts is a great...great way to include researchers from different locations. Having virtual face-to-face meetings as opposed... collaboration includes scientists working in different time zones, it’s important that everyone can access...project at the International Cancer Immunotherapy Conference in Frankfurt, Germany When funding and circumstances...techniques are being performed the same way at different sites. I did a lab exchange as part of the collaboration...been able to do from my home lab. Scientific conferences and workshops are another great place to meet...

  8. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    Published
    June 30, 2015, 2 p.m.
    ...system which has been designed to perform gene transfer in vertebrates. The system is comprised of 2 components...transposase has been discovered in the genomes of many different fish species in a variety of non-functional forms...the lab showed that efficiencies of stable gene transfer using SB100x were comparable to stable transduction... vectors were developed into 16 variants with different selection markers (blasticidin, neomycin, puromycin... each contain two SfiI restriction sites with different overhangs depending upon whether they are on the...only likely to grow in the coming decades.  References 1. Kowarz E, et al. Optimized Sleeping Beauty... Beauty transposase enables robust stable gene transfer in vertebrates. Nat Genet. 2009 Jun;41(6):753-...

  9. Plasmids 101: Origin of Replication

    Type
    Blog Post
    Published
    Nov. 11, 2020, 2:23 p.m.
    ...cells. The replicons of plasmids are generally different from the those used to replicate the host's chromosomal...include ColE1, pMB1 (which comes in a few slightly different but well known derivatives), pSC101, R6K, and ...regulated. Generally, control of replication is referred to as "relaxed" or "stringent" depending on whether...maintains about 20 copies per cell, while pUC – which differs by only two mutations – will produce as many as...considerations. *Requires pir gene for replication (reference). **F1 is a phage-derived ori that allows for...particles. Plasmids with phage-derived ori's are referred to as phagemids.   Other factors that affect...clarified? If so please, post in the comments.  References Chatterjee A, Sun SB, Furman JL, Xiao H, Schultz...

  10. Viral Vectors 101: What is a Virus (and a Viral Vector)?

    Type
    Blog Post
    Published
    Aug. 18, 2023, 4:15 p.m.
    ...out if you want to learn more. References and resources References Balloux, F., & van Dorp, L. (2017...called an envelope. Viruses without an envelope are referred to as naked or non-enveloped, while viruses with...viruses, here’s a quick tip on terminology. “Virus” refers to a single type of virus, not the individual viral...infection — researchers were able to create far safer viral vectors that can be used in BSL-1 or BSL-2...experimentally controlled conditions, making them far safer to work with than virions. The four types of viral...vectors (which many researchers will informally refer to as viruses) are named after the viruses they’...

  11. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    Published
    April 7, 2015, 4:21 p.m.
    ...of many complex cellular behaviors, such as differentiation or metabolism. Our attempts to manipulate these...cerevisiae with few techniques that are easily transferrable across host species or metabolic pathways. A...system for multidirectional control The CRISPR interference (CRISPRi) model consists of a catalytically .... cerevisiae by using three RNA programs to differentially alter the levels of five genes that encode ...sequentially rerouted the metabolic flux to generate differently pigmented derivatives of violacein. Using dCas9...that will still recognize guide RNA but with a different dCas9 binding module, alternative sets of transcriptional...pharmaceuticals. Follow him on Twitter @AdamChinFatt     References Qi, L. S. et al. Repurposing CRISPR as an RNA-guided...

  12. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    Published
    June 24, 2020, 5:45 p.m.
    ...activation p300 acetyltransferase dCas9 fused to the catalytic domain of p300 acetyltransferase increases levels...transcription, also includes light-regulated histone methyltransferases and deacetylases. CRISPR and epigenetics ...delivery systems. Transcriptional repression DNA Methyltransferase 3 Alpha (DNMT3A) Vlatka Zoldoš’ lab has deposited...repression. Created with BioRender.com. DNA Methyltransferase MQ1 Margaret Goodell's lab has deposited pcDNA3.1...Q147L)-EGFP, a fusion of dCas9 to a small DNA methyltransferase from the prokaryote Mollicutes spiroplasma...Schwiesow contributed to updating this article. References Haldeman JM, Conway AE, Arlotto ME, et al (2018... Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers...

  13. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    Published
    Feb. 8, 2024, 2:15 p.m.
    ...they do arise. Happy CRISPR’ing! References and Resources   References Rabinowitz R, Offen D. Single-Base...cutting to occur. The RNA sequence and PAM site confer specificity of the nuclease; however, many Cas ...effects should impact your experimental design differently. Yes, it is always better to reduce off-target...mutation, off-targets must be quantified slightly differently. For a comprehensive approach, RNA-seq would ...to control for off targets as well as clonal differences. If you are performing CRISPR library screens... as a transfection or transduction control, a different gRNA would have distinct off-target sites, eliminating... there really aren’t any effective off-target reference controls. Off-targets and your experiment Every...

  14. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    Published
    Dec. 20, 2022, 2:15 p.m.
    ...by Hannah Dotson in Dec. 2022.  References and Resources References Gossen M, Bujard H. Tight control...CMV, EF1A, and SV40, are always active and thus referred to as constitutive promoters. Others are only ...In the presence of tetracycline (Tet), TetR preferentially binds Tet over the TetO elements, allowing ...hear some transcription factor and promoter pairs referred to as binary systems. Binary systems are genetic... One can therefore place GAL4 and GAL80 under different promoters and create sophisticated patterns of...wondering, “why is it necessary to have all 3 of the different binary systems above if they all behave very similarly...s00253-018-9485-4. Epub 2018 Nov 3. PMID: 30392122. Pfeiffer, Barret D., et al. “Refinement of Tools for Targeted...

  15. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    Published
    July 28, 2016, 2:30 p.m.
    ...For more context, exposition, and detail, please refer to the paper. Why might one be interested in state...register (Fig. 1C below). If attB-attP pairs from different recombinases are overlapping or nested on a register...that enters distinct states when treated with different combinations and orders of two recombinases. ...leveraging qPCR data from multiple primer pairs, one can infer the predominant state of registers within a sample...strong, directional terminators and promoters with buffer sequences on their 5’ and 3’ ends (for insulation... register. The purpose of this is to prevent interference between the transcribed gene and its 5’ UTR.... in biocomputing and biological circuits.     References 1. N. Roquet et al., Synthetic recombinase-based...

  16. Quantifying DNA? Here are Five DNA Quantification Methods to Consider

    Type
    Blog Post
    Published
    July 21, 2020, 1:15 p.m.
    ...the liquid. Molecules absorb different wavelengths of light to different degrees and many molecules have...cuvettes. First you measure the absorbance of the buffer that the DNA is in. This is a “blank” and measures...the ladder. You then run samples of your DNA at different dilutions and quantify based on band intensities...elute and store your plasmid DNA Learn about the different grades of plasmid DNA preps Resources on Addgene.org...Addgene.org Visit the Molecular Biology Reference Find plasmids for your research Browse protocols for ... Jennifer Tsang...

  17. Plasmids 101: Gram Negative and Positive Bacteria

    Type
    Blog Post
    Published
    Aug. 2, 2022, 1:15 p.m.
    ...in 1983. The differences between gram positive and negative bacteria is due to differences in cell wall...incredibly helpful in visualizing differentiations between bacteria, leading to safer medicine and more successful...minoring in studio arts photography.  References and resources References Silhavy, TJ., Kahne, D. & Walker ...negative and positive bacteria are permeable to different classes of antibiotics. The classification system...

  18. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    Published
    Oct. 19, 2023, 1:15 p.m.
    ... 2019), and different combinations of ITR and capsid serotypes vary in their gene transfer efficiency....DNA molecule as the gene that they package. In a transfer plasmid, the transgene is flanked by two ITRs,...vectors, modifications can drastically affect gene transfer efficiency and transgene expression (Zhou et al...expression can be altered by using ITR sequences from different AAV serotypes that vary in their RBE and T-shaped...restriction digest methods.    Resources and references More resources on the Addgene blog Viral Vectors...Addgene.org Addgene's AVV guide AAV Viral Preps References  Earley, L. F., Conatser, L. M., Lue, V. M., ...

  19. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ... the transcript. Different Cas orthologues and subtypes function somewhat differently and can be more ...Transcriptional repression (CRISPRi) in bacteria As RNA interference does not function in bacteria, most efforts ...promoters or direct gene knockout. In contrast, CRISPR offers a much more user-friendly way to modulate gene ...Peters, Oren Rosenberg, and Carol Gross used two different genetic systems, targeting either Gram-negative...bacterial species (Peters et al., 2019). The systems differ on how exactly the CRISPRi locus is introduced ...Clostridium difficile, could be targeted and preferentially depleted from gut. While still far from the...the BE3 that further improves editing rates. References Bikard D, Euler CW, Jiang W, Nussenzweig PM, ...

  20. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    Published
    Feb. 13, 2025, 2:15 p.m.
    ...updated by Jennifer Tsang in August 2020 and by Emily P. Bentley in February 2025. References and Resources...for resolving DNA double-strand breaks (DSBs) preferred by researchers. Compared to non-homologous end...APOBEC3G displayed less predictable sequence preferences.  The Liu lab also used natural and engineered...generation of base editors. Compared to BE3, BE4 offers a 2.3 fold decrease in C→G and C→A products as ...yielded an editor with more flexible sequence preferences, evoAPOBEC1-BE4max, which is significantly more...the development of prime editing, which uses a different mechanism than base editing, has allowed scientists...Resources References Chen, L., Zhu, B., Ru, G., Meng, H., Yan, Y., Hong, M., Zhang, D., Luan, C., Zhang, ...
Showing: 241 - 260 of 867 results