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We narrowed to 407 results for: expression vector

Showing: 221 - 240 of 407 results

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

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    Blog Post
    ...transgene. Typically, the desired target of RNAi is expressed as inverted repeats separated by a loop region...considerations, such as the starting material (DNA-based vector or purified RNA) and the initial state of the dsRNA... easiest approach to induce RNAi involves the expression of long hairpin RNA (hpRNA). This method is facilitated...construct has unique features, such as inducible expression of the hpRNA and/or fusing your gene target sequence... Although downstream work is required, several vectors can be found at Addgene that could serve as an ...an excellent start (#22988, #137884). Both vectors allow the creation of amiRNAs based upon miRNA precursors...

  2. Genome-wide Screening Using CRISPR

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    Blog Post
    ... deliver a pooled population of gRNAs to Cas9 expressing cells. Pooled lentiviral CRISPR libraries (referred...population of gRNA-containing lentiviral transfer vectors, each targeting a specific gene within the genome...gRNAs are then cloned into lentiviral transfer vectors to create the CRISPR library. CRISPR libraries ...) to be used to generate lentivirus (B). Cas9-expressing cells or wild-type cells are treated with lentivirus...

  3. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

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    Blog Post
    ...in a single vector. In addition, the pol III promoter is relatively weak, and low expression of gRNAs from...transcription, potentially increasing the levels of gRNA expression. Since tRNAs are among the most abundant molecules... confirming gRNA cleavage, they examined gRNA expression levels and indel generation for two PTG constructs...protoplasts. Compared to matching traditional gRNA-expressing constructs, transcript levels of PTG1 and PTG2...Xie et. al are specifically designed for plant expression, PTGs will likely be adaptable to many different...capacity. Still confused about the various tRNA expression strategies? No worries, I've summarized the three...research? Check out the PTG plasmids or plant expression systems available from Addgene. If you have any...

  4. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    ...enzyme cut sites Open reading frame finder  Addgene vector database: Find sequences for plasmids assembled... sequence and functional information, such as expression patterns, post-translational modifications and...repository for validation data on Addgene/CLOVER viral vectors.  Caspedia: A data repository for Cas enzymes.  ...

  5. Hot Plasmids and Viral Preps - September 2021

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    Blog Post
    ...of robust expression of high G+C content genes, as validated through monitoring the expression of fluorescent...different vectors and flow cytometry is used to sort populations by mCherry and GFP expression to check... be seen through the success of the change in expression of GFP, for the stop codon should have changed...

  6. K. phaffii: Rising to the Occasion

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    Blog Post
    ...Phillips made the decision to release the K. phaffii expression system to the general research community in the...different uses for the phaffii-lous strain. Protein expression   K. phaffii first gained attention for its powerful...This gene promoter, AOX1, was co-opted for the expression of recombinant proteins by the pharmaceutical...pharmaceutical and biotech industries as a protein expression workhorse. This yeast can also give more protein bang...Saccharomyces cerevisiae to human: The important gene co‑expression modules. Biomedical Reports 7.2 (2017): 153-158...Five Popular Model Organisms Plasmids 101: Yeast Vectors Tips for Screening with the Yeast Two Hybrid ...

  7. Tips for Using FRET in Your Experiments

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    Blog Post
    ...interest: Plasmid Color Expression Description pPROEX Aqua Cyan Bacterial Expresses Aquamarine with N-terminal...Cyan Mammalian Expresses mammalian optimized Aquamarine mCerulean N1 Cyan Mammalian Express a gene of interest...Cyan Mammalian Expresses mammalian optimized CyPet pCyPet-His Cyan Bacterial Expresses CyPet with C-terminal... Mammalian Expresses mammalian optimized YPet pYPet-His Yellow Bacterial Expresses YPet with C-terminal...Mammalian Express a gene of interest fused to the C-terminus of SYFP2 Clover Green Mammalian Expresses Clover...Cyan & Yellow Bacterial Gateway-compatible vector to express a gene of interest fused between ECFP and ...monomeric Cerulean mCerulean C1 Cyan Mammalian Express a gene of interest fused to the C-terminus of monomeric...

  8. Hot Plasmids - October 2022

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    Blog Post
    ... available as Cre-dependent, EF1ɑ-driven viral vectors (AAV1) - and you can also find several other constructs...B) Cortical slice of HcKCR1-EYFP and tdTomato expressed layer 2/3 neurons in mouse. C) Action potential...voltage traces of action potentials in a HcKCR1-expressing mouse neuron without (top) and with (bottom) ... blot for the myc-tagged protein GFP-smFP-myc expressed from Plasmid 98926. See the Applications section...

  9. Plasmids 101: Using Transposons in the Lab

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    Blog Post
    ...in viral vectors, but easily fit in a TE. Transposons are also less likely than viral vectors to induce...Non-autonomous transposons, however, need another TE to express a transposase or reverse transcriptase so they ...transposon’s LTRs, similar to when packaging viral vectors.   Let’s take a closer look at three different ... in human cells at rates similar to retroviral vectors. The hyperactive version of the SB transposase,...efficiency in human cells similar to retroviral vectors. The hyperactive PB transposase (hyPB) is ~10-fold...transfer Transposons are an alternative to viral vector delivery of transgenes, like for iPSC reprogramming... piggyBac, which is a big advantage over viral vectors (~5 kB payload for AAV and ~8 kB payload for lentivirus...

  10. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...LinkedIn.   References 1. Yin, R., et al. "Gene expression profiling analysis of bisphenol A-induced perturbation...cytoskeletal disorganization, apoptosis, and gene expression disregulation on HeLa cells." Journal of applied...with the Agarose Method Plasmids 101: Mammalian Vectors Resources on Addgene.org Find mammalian plasmids...plasmids for your research Find viral vectors for infecting mammalian cells ...

  11. CRISPR-mediated Plant Base Editors

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    Blog Post
    ...plant We developed plant adenine base editor (ABE) vectors to install A>G or T>C mutations in the plant genome...genome. We constructed three versions of the vector (pKABE6.3, pKABE7.9, and pKABE7.10) so that the user...are better suited (Gaudelli et al., 2017). Our vectors have two Bsa1 sites to clone the single guide RNA...scaffold) in a pol-III (OsU3) promoter driven expression system. We use a single polycistronic tRNA-gRNA... genomic site simultaneously.     These binary vectors could be used to generate stable genome edited ...Agrobacterium-mediated transformation. We also constructed smaller vectors suitable for transient assays using protoplast ...

  12. Plasmids 101: The Wide World of Natural Plasmids

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    Blog Post
    ...other molecules to host organisms. For instance, vectors derived from the Agrobacterium Ti virulence plasmid...especially when combined with natural inducible expression systems that allow tuning of metabolic pathways...developing new cloning techniques based on these vectors (Shareck et al., 2004). Plasmid categorization..., Y., Lee, B., & Miguez, C. B. (2004). Cloning Vectors Based on Cryptic Plasmids Isolated from Lactic ...

  13. Plasmids 101: Origin of Replication

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    Blog Post
    ...plasmids with low to medium copy numbers can still express massive amounts of protein given the proper promoter...the table below for more details.   Common Vectors Copy Number+ ori Incompatibility Group Control ...Relaxed This table defines common cloning vectors, their copy number, ori, and incompatibility groups...

  14. Plasmids 101: How to Verify Your Plasmid Using a Restriction Digest Analysis

    Type
    Blog Post
    ... Congratulations, you have a plasmid expressing your gene of interest (YGOI) and are ready to dive into...confirm the presence of an insert in a particular vector by excising the insert from the backbone. To do...will need to know both the approximate size of the vector backbone as well as the predicted size of the insert...

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

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    Blog Post
    ...My lab's vector of choice is AAV, with nearly every experiment requiring AAV. Before joining my lab, ...shell, or capsid; 2) the pHelper plasmid which expresses adenovirus genes which help mediate AAV viral ...Production of recombinant adeno-associated viral vectors for in vitro and in vivo use. Current protocols... out what makes AAV a versatile tool for gene expression in mammals Use the PEI calculator for planning...

  16. Plasmids 101: Protein tags

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    Blog Post
    ...Fluorescent Protein (GFP) Fluorescent Protein Guide Expression Vectors from the Berkeley QB3 MacroLab Eric thanks...box of tags and fusion proteins to aid in the expression and purification of recombinant proteins. Tags...purification to get your protein as pure as possible? Vectors that offer different combinations of tags are readily...Biochemists and molecular biologists who need to overexpress and purify proteins can face any number of technical... some of the hurdles to overcome in order to overexpress a recombinant protein? It is not generally in...in a cell’s best interest to overexpress a protein. Energy and cellular resources are being spent to make...stabilization, as can maltose-binding protein (MBP). Overexpression can also lead to insolubility, and aggregated...

  17. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

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    Blog Post
    ...in patched cells expressing wild type CoChR, these APs are absent in soCoChR-expressing cells. Finally,...C-terminus of the protein. Unlike the diffuse neuronal expression observed with wild type CoChR, soCoChR localizes...stimulation are largely absent in the soCoChR-expressing system. Taken together, Shemesh et. al. describe...Pseudomonas putida CBB5 and has been tailored for expression in E. coli. This operon is under the transcriptional...strong constitutive promoter (BBa_J23100) and expresses N-demethylases and other required enzymes, including...Moreover, a caffeine “addicted” E. coli strain expressing this operon can serve as a living biosensor for...FRET reference page for many other common FRET vectors. Murakoshi H, et al. Sci Rep. 2015. Pubmed PMID...

  18. PEI Calculator for Planning AAV Packaging Transfections

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    Blog Post
    ...Production of Recombinant Adeno‐Associated Viral Vectors for In Vitro and In Vivo Use." Current Protocols...When Using AAVs AAVs Are Great Tools for Gene Expression in Mammals Resources on Addgene.org Find AAV...

  20. Neuronal labeling with Spaghetti Monster

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    Blog Post
    ...epitope tags, low FP expression may be insufficient for monitoring a weakly expressed POI. Spaghetti Monster...cytotoxicity, low immunogenicity, and the long-term expression of AAV delivered genes. AAV serotypes have been.... Thanks to this work it is now much easier to express molecules in your favorite neuronal population ...insufficient for detection when the POI is weakly expressed. As these epitopes tags have no intrinsic visual... be fused to scaffold proteins in order to be expressed stably in neurons. As compared to epitopes tags...labeling of neurons. As discussed above, weakly expressed proteins in particular are difficult to track ...other). Combined with AAVs, FPs can be easily expressed in the brain and can be detected through their...
Showing: 221 - 240 of 407 results