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We narrowed to 405 results for: expression vector

Showing: 221 - 240 of 405 results
  1. Mouse Modeling, Part 2: Breeding and Crossing Mice

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    ... that expresses Cre with a strain that expresses a floxed gene of interest. Cre may be expressed globally...the appropriate primers to detect your targeting vector or gene of interest. 2. Breed your parental mice...infertility and can arise due to copy number, expression level, and site of integration of the transgene...in cell lines to determine how it impacts the expression level of the transgene. In existing strains of...heterozygous background to reduce copy number or expression level effects.  Another challenge that may arise...heterozygous for each mutated allele.  Cre-lox expressing strains Cre-lox strains are a common example ...we explained how genetic recombination in mice expressing both Cre and LoxP occurs. But how are these mice...
  2. Hot Plasmids - February 2022

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    ...individually expressed proteins run in separate lanes. B: individually expressed and coexpressed proteins ...through two polycistronic expression vectors (3570 lanes/50 mLs) coexpressing a set of either 10, 30, 50...editing within a single expression cassette is advantageous to Cas9’s larger expression requirements (Figure... Figure 2: comparison of Cas 9 vs. Cas 12a expression cassettes when encoding multiple guides. While...opportunity for lab members or students new to protein expression and purification. Figure 3: The Penn State...Santilli, et al. 2021 Plasmids are used to express 10, 15, 20, 30, 40, 50, 60, 80 and 100 kD proteins...
  3. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

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    ...then easy to package in low-capacity vectors like AAV, a popular vector due to its low immunogenicity. But...in size and are easy to package in low-capacity vectors! These small, but mighty type VI-D enzymes are ...collateral cleavage does not occur when Cas13d is expressed in a mammalian system. Since Cas13d is functionally...
  4. RNA Interference in Plant Biology: New Tools for an Old Favorite

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    ...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...
  5. Common Injection Routes in Mice

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    ...be injected, including cells, drugs, DNA, viral vectors, and viruses. All researchers must go through ...choice for administering different drugs and viral vectors. Like IV injections, intraperitoneal injections...for the delivery of various therapies and viral vectors. This injection sounds very disruptive, but surprisingly... This is most often used for delivery of viral vectors, though drug treatments are also possible. One ...Addgene.org Genome Engineering Guide CRISPR Mammalian Expression plasmids Cre-Lox plasmids References Prathipati...
  6. Genome-wide Screening Using CRISPR

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    ... 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...
  7. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

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    ...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...
  8. Hot Plasmids and Viral Preps - September 2021

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    ...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...
  9. K. phaffii: Rising to the Occasion

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    ...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 ...
  10. Tips for Using FRET in Your Experiments

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

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    ...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.  ...
  12. Fluorescent Proteins 101: Luciferases

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    ...gene expression. The regulatory element is cloned upstream of the luciferase gene, and expression of luciferase...backbones with the luciferase gene, a variety of expression vectors, and literally thousands of reporter constructs...wavelength of light they emit or to optimize their expression in mammalian systems. Our luciferase collection...luciferase assays have become a gold standard in gene expression analysis to investigate the effect of regulatory...interactions Split luciferases, such as NanoBiT, are expressed in two subunits and only catalyze a luminescent...
  13. A Tour of Addgene's Most Popular Pooled Libraries

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    ... genes, creating mutations that prevent their expression. The knockout cells can be used to identify genes...Library, whose acronym means Modular, NGS-trACKable ExpRession Element. This library consists of broad host ...interest are fused to anchoring proteins to be expressed on the surface of a host cell or virus, such as...Additional resources on the Addgene blog Viral Vectors 101: Preparing Pooled Libraries Pooled Library ... MORF library, which stands for Multiplexed Overexpression of Regulatory Factors. The Feng Zhang lab developed...
  14. Hot Plasmids - October 2022

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    ... 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...
  15. 10 Basic tips for mammalian cell culture

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    ...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 ...
  16. Plasmids 101: Using Transposons in the Lab

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    ...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...
  17. CRISPR-mediated Plant Base Editors

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    ...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 ...
  18. Plasmids 101: The Wide World of Natural Plasmids

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    ...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 ...
  19. Plasmids 101: Origin of Replication

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

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    ... 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...
Showing: 221 - 240 of 405 results