Skip to main content
Addgene
Open Menu

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

Information

In Category

We narrowed to 591 results for: cel.2

Showing: 41 - 60 of 591 results

  1. Adeno-associated virus (AAV) Plasmids

    Type
    Collection
    ...should be handled at BSL-2. AAV can transduce both dividing and non-dividing cells with a low immune response...expressing Rep2 and Cap1 James M. Wilson 232199 pAAV2/2 AAV2 AAV packaging plasmid, expressing Rep2 and Cap2...e.g., pHelper) and specific packaging cell lines (e.g., HEK293 cells) during AAV production. Read our AAV...

  2. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    Blog Post
    ...the start codon of retinal cell-type specific genes from mice. Strategy #2 (ProB): Create promoters using...horizontal cells transduced by both AAVs would express Cre-mCherry and GFP.   Figure 2: Synthetic...doi.org/10.1038/s41593-019-0431-2  Neff, E.P. An AAV library for retinal cell expression in mouse, macaque... horizontal cells, and Cre-dependent-GFP expresses in horizontal cells and ganglion cells, only the horizontal... in retinal cells. To do this, the group developed a new strategy for finding retinal cell-type specific...and high cell-type specificity.  Strategy #3 (ProC): Create promoters by repeating mouse cell type specific... to test for cell type-specific expression of GFP. Half (113) expressed in retinal cells four weeks post-injection...

  3. Zinc Finger Consortium: Zinc Finger Arrays

    Type
    Collection
    ...gene modification. Maeder ML et al. Mol Cell. . 2008. Jul 25. 31(2):294-301. PubMed PMID 18657511 . Oligomerized...OPEN OPEN gGTCATCTGCcattcGAGGCGGACa REST corepressor 2 OZ553 and OZ554 OPEN OPEN gCTCACCATCtgttggaTGCGATGGAa...

  4. Cellosaurus: A Cell Line Information Database

    Type
    Blog Post
    ...can browse entries by cell line groups, such as their vaccine production, SARS-CoV-2 research, or adenovirus...Knowledge Resource. J Biomol Tech. 2018; 29(2): 25-38....in which the cell line was described. Using Cellosaurus Cellosaurus is searchable by cell line name (recommended...resource called Cellosaurus, a growing cell line information database describing nearly 145,000 cell lines. Cellosaurus... Cellosaurus Cellosaurus seeks to identify and describe all cell lines used in research, using four methods... related cell lines when annotating a cell line Community submissions via email: cellosaurus@sib.swiss...with cell line catalogs and collections. Importantly, Cellosaurus also identifies problematic cell lines...

  5. CRISPR Guide

    Type
    Collection
    ...Haploid Embryonic Stem Cells Carrying a Guide RNA Library. Cell Stem Cell . 17 (2), 221–232. PMID: 26165924...used to transduce your experimental cell population (Figure 8E). In a 2-vector system, you’ll need to either...of CRISPR-Cas9 with Bacteriophage Proteins. Cell , 168 (1–2), 150-158.e10. PMID: 28041849 Sakuma, T., Nishikawa... regulation of transcription in eukaryotes. Cell . 154 (2), 442–451. PMID: 23849981 Gilbert, L. A., Horlbeck...Transcriptional Programs with CRISPR RNA Scaffolds. Cell . 160 (1–2), 339–350. PMID: 25533786 Cas9 alternatives...single RNA-Guided endonuclease of a Class 2 CRISPR-CAS system. Cell , 163 (3), 759–771. PMID: 26422227 Zetsche...left free to interact with the target DNA. Figure 2: Overview of the NHEJ repair mechanism Cas9 will only...

  6. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    ...Disable the other repair mechanisms Mammalian cells have 2 primary repair mechanisms besides HR - Non-homologous... However, if you have a larger knock-in (200 bp – 2 kb), a dsDNA donor will need to be used due to synthesis...accidentally change your protein of interest.   Fig. 2: Stop re-cutting with PAM and guide RNA disrupting.... Com., 8(2024), (2017). 10.1038/s41467-017-01836-2 Charpentier, M., Khedher, A. H. Y., Menerot, S., Brion...varying across cell types. Variance can be caused by cell cycle rates; rapidly diving cells will have higher... of your cells. Enrich S-phase cells HR operates predominantly in S and G2 phase of the cell cycle, when...HDR events, ensure cells are actively cycling through the cell cycle stages. If cells are overly confluent...

  7. AAV Production in HEK293 Cells

    Type
    Protocol
    ...3319, 3180 cm 2 Cellstack 2, Corning 3269, 1272 cm 2 Heat-inactivated FBS (HI-FBS) Pro-Tip Different brands... 4 °C. Cell Lysis Buffer : 50 mM Tris HCl, 150 mM NaCl, 2 mM MgCl 2 Add the following to the 2 L sterile... solution can be stored at 4 °C for up to 2 months. After 2 months, discard the tube and thaw a new working... mL of PBS. Aspirate PBS and add 2 mL of 0.05% Trypsin/EDTA. Wait ~2 min. Neutralize trypsin by adding...Seed cells in CS2 Day 2: Seed cells in CS5 Day 3 (am): Transfect cells Day 7 (am): Harvest cells Equipment...times to obtain a single cell suspension (no clumps of cells). Pool cells from 2 x T-175 flasks. Adjust ...touching the cells when replacing media. T-175 flask, Corning 430825, 175 cm 2 Cellstack 5, Corning 3319...

  8. Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification

    Type
    Blog Post
    ...non-specific proteins, and then elute YFP.  Option #2: Opto-Nanobodies (OptoNBs) purification You skip adding...blue lights. Which option do you choose?  Option #2 may sound like fiction, but light-controlled protein...experiments.  AsLOV2 Inertion Site mCherry #1 mCherry #2 GFP Loop 1 dark-induced dark-induced dark-induced...OptoNBs to control cell signaling The second application for OptoNBs takes us into cells: light-based control... regulation of endogenous signaling activity in cells.  Let’s shed some light on the key components of...their use for protein purification and controlling cell signaling. Key components of OptoNBs OptoNBs have...can bind, it co-localizes with its target at the cell membrane and when the OptoNB can’t bind, it localizes...

  9. Photosensitizer Induced Cell Ablation with FAP-TAP MG-2I-dL5**

    Type
    Blog Post
    ... post treatment (see figure 2). Induced cell death was confirmed by TUNEL staining.  The video below shows...fluorogen activating protein - dL5** in this case), and 2) a fluorogen (the TAP - targeted and activated photosensitizer...PMID: 26808669. PubMed Central PMCID: PMC4916159. 2. Jacobson, K., Rajfur, Z., Vitriol, E., & Hahn, K....HEK293 cell culture and in live zebrafish. For cell culture experiments, only dL5**-expressing cells treated...inactivate target proteins or cells while having little effect on neighboring cells. The photosensitizer used...Here Light-induced cell ablation with MG-2I-dL5** He et al tested out light-induced cell ablation with MG...effective killing of dL5** cells without death of neighboring bystander wild-type cells.   Zebrafish experiments...

  10. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

    Type
    Blog Post
    ...Induction of Human Germ Cell Fate from Pluripotent Stem Cells. Cell Stem Cell, 2015. 17(2): p. 178-94. PubMed... of human induced pluripotent stem cells. Stem Cell Rev, 2015. 11(2): p. 242-53. PubMed PMID: 25578634...differentiated somatic cell to revert back to a pluripotent stage (iPSC formation) [2, 3]. The iPSCs then...somatic cells via various methods. Some of the more common delivery methods include retrovirus [2], lentivirus...PubMed PMID: 16810240. 2. Takahashi, K., et al., Induction of pluripotent stem cells from adult human fibroblasts...three routes to regeneration. Nat Rev Mol Cell Biol, 2011. 12(2): p. 79-89. PubMed PMID: 21252997. 37. Merrell...differentiation of human ES cells and iPS cells into mature pancreatic insulin-producing cells. Cell Res, 2009. 19(...

  11. Plan Your Experiment

    Type
    Collection
    ...your target cells. There are a variety of ways to deliver and express these components (Figure 2), and the...optimal system will likely depend on your cell type. Figure 2: Different delivery systems utilized for ...and/or Cas9 expression or (2) a lack of efficient target cleavage in cells that do express both Cas9 and...CRISPR-Based genome editing. Trends in Biotechnology , 36 (2), 173–185. https://doi.org/10.1016/j.tibtech.2017.11.006...difficult-to-transfect cells. You can direct expression to specific cell types by using cell-specific promoters...human embryonic kidney 293 (HEK293) cells. For more difficult cell types, you may need to look into other...are good choices for more difficult cell types like primary cells. For CRISPR delivery, lentiviral and...

  12. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ...postmitotic, long-lived cell types. AAVs in the clinic Thus far, AAV vectors AAV1, AAV-2, AAV1-AAV2 hybrids...neutralizing antibodies against some AAV types, AAV1, 2, 3 and 5 are thought to be as high as 70% among the... 10.24 (2003): 1999-2004. PubMed PMID: 14566358.  2. Hirsch, M. L., et al. "Self-complementary AAV mediates... do so, their T-cells were grown in vitro, their mutations corrected, and the T-cells were transferred...functioning T cell repertoire (1). In this example, it was possible to transduce the T-cells outside the...a wide range of cell types including but not limited to muscle, liver, and brain cells. There are a multitude...chain (c) and is characterized by a lack of immune cell development and function. To date, gene therapy ...

  13. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ...splitting transfected cells into 3 plates at different densities for selection). A 2% agarose solution is...transition from a 48-well cell plate to a 96-well assay plate. We generally freeze at least 2 ampules each of 4...of agarose, which is then overlaid with the firmer 2% agarose gel solution discussed above for picking.... a wide variety of cell lines, from HEK to CHO cells to neural and endocrine cells such as AtT-20, PC12...then handled. Additionally, many cell lines, especially the endocrine cell lines we most commonly work with...specific cell line, and its transfection efficiency. We often trypsinize the transfected cells on the day...increased). For some cells, add a drop of serum to aid in outgrowth. Depending on the cell type, we sometimes...

  14. Antibody Guide

    Type
    Collection
    ...surface of most immature B cells; function is unknown Monomer with a valency of 2 IgE Protects against parasites...such as anaphylactic shock Monomer with a valency of 2 IgG The most abundant isotype in blood, also found...secondary immune response Monomer with a valency of 2 IgG1; IgG2; IgG3; IgG4 IgM Produced during the primary... ranging from antibody fragments (shown in Figure 2) to completely unrelated molecules. Examples include...used for assembling protein nanostructures. Figure 2: An antibody (IgG, far left) compared to alternative... produced. Cells - Antibodies can also be produced in cell cultures using hybridomas, cell cultures created...created by fusing B cells with myeloma cells (immortalized B cells). The genetic reorganization that creates...

  15. Zinc Finger Consortium: Nuclease Expression Vectors

    Type
    Collection
    ..., Nat Biotech 2007 ) in mammalian cells or zebrafish). pMG10 (a '“2-in-1” ZFN cassette for expression ...vectors designed to express nucleases in different cell types. Zinc finger arrays can be transferred in-...vectors designed to express nucleases in different cell types. Zinc finger arrays created using ZFC reagents...nuclease domains: pST1374 (for expression in mammalian cells or zebrafish) pDW1775 (for expression in plants)... Miller et al., Nat Biotech 2007 ) in mammalian cells or zebrafish (can be used as a substitute for pMLM335... Miller et al., Nat Biotech 2007 ) in mammalian cells or zebrafish (can be used as a substitute for pMLM336... Miller et al., Nat Biotech 2007 ) in mammalian cells or zebrafish). pMLM802 (for expression of a ZFN ...

  16. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...AAV); 8–10 kb (lenti) 38 kb Cells HEK-293T Sf9 Time to collection 2–4 days 5–10 days Purification... R.M. (2002). Insect cells as a factory to produce adeno-associated virus type 2 vectors. Human Gene Therapy...Centrifugation Titration time ~1 day (AAV); ~1 week (lenti) 2–10 days Initial titers Higher Lower Resources...that already do mammalian cell culture. HEK-293T cells are the most common cells used in viral production...versus insect cells for viral production Consideration Mammalian cells Insect cells Viruses AAV ...type of cells that will be used to make the virus. Viruses can be produced in many different cell types,...produced in bacteria cells. However, some viruses can be produced in multiple different cell types, including...

  17. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

    Type
    Blog Post
    ...deletions in human cells using zinc finger nucleases. Genome Res. 2010 Jan;20(1):81-89. PubMed. 2. Bauer DE, ...with TALENs, in collaboration with the Porteus lab [2]. With the advent of CRISPR/Cas9, we began to explore...CRISPR plasmids into MEL cells by electroporation, sort your transfected cells via FACS, primer validation...article "Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9" from the Journal of Visualized...CRISPR/Cas9 to create genomic deletions in mammalian cell lines. Below Bauer and Canver discuss the motivations...deletions is to produce monoallelic deletion clones. Cells with just a single intact copy at a given locus ...screening by conventional PCR. Biallelically disrupted cells can be easily identified. Even in the case of clones...

  18. CRISPR Plasmids - Tagging

    Type
    Collection
    ...gene-specific CRISPR and donor vectors as well as pX330S-2-PITCh, expressing a generic gRNA targeting the PITCh...Protocols, 2016 . Yamamoto Tagging Plasmids: pX330S-2-PITCh - expresses Cas9 & PITCh-gRNA pX330A-FBL/PITCh...Drosophila cells. N terminal tagging in Drosophila cells 3.2 MB C terminal tagging in Drosophila cells 3.3 MB...following article: Natsume, et al. Cell Reports 2016 Förstemann Drosophila Cell Tagging System The Förstemann... Allen Institute for Cell Science Plasmid Collection The Allen Institute for Cell Science has produced...tagged, human, induced pluripotent stem cell (hiPSC) lines. These cell lines were created using CRISPR and...tagging transcription factors with FLAG in human cell lines. To repeat the tagging in your system, use...

  19. AAV Viral Preps

    Type
    Collection
    ...AAV vectors for trial purposes in serotypes AAV1, 2, 5, 6, 8, 9, 11, and retrograde Retrograde AAV To ... tools that effectively transduce many mammalian cell types. AAV is used in a wide range of experiments...Tetracycline Transactivators, Affinity Purification, Cell Ablation, CRISPR Monosynaptic Neuronal Tracing Rabies...

  20. Optogenetics Plasmid Collection

    Type
    Collection
    ...optogenetic tool is the widely used Channelrhodopsin-2 (ChR2) from Chlamydomonas reinhardtii . ChR2 is a ...switch tools to activate, inhibit, and manipulate cellular processes. Plasmid...to blue light and allows cations to flow into the cell. Variants of channelrhodopsin have been engineered...improve the expression and membrane targeting in host cells. Other microbial opsins, such as the Halorhodopsins...actively move cations or anions into or out of the cell. When using these opsins as a tool to control neuronal...plasmids that use light to control a protein or cellular activity. For example, search "Cdc42", "transcription...
Showing: 41 - 60 of 591 results