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  1. CRISPR History and Development for Genome Engineering

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    ...RNA-guided regulation of transcription in eukaryotes. Cell . 154(2):442-51. PMID: 23849981 Ishino Y, Shinagawa ...Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell . 163(3):759-71. PMID: 26422227 Zetsche...transcribed to make the pre-CRISPR RNA (pre-crRNA). (2) The pre-crRNA is processed into individual crRNAs...2015. Class 1 (Multi-subunit effector complex) Class 2 (Single multi-domain effector) Type I (Cas3) Type ... with unprecedented speed and specificity. Figure 2: An overview of CRISPR and NHEJ/HDR. The Cas9/gRNA...genomic loci in living human cells by an optimized CRISPR/Cas system. Cell . 155(7):1479-91. PMID: 24360272... for Mapping Multiprotein Complexes in Human Cells. Cell Rep . 13(3):621-33. PMID: 26456817 Fu Y, Sander...
  2. 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...
  3. Photosensitizer Induced Cell Ablation with FAP-TAP MG-2I-dL5**

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    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...
  4. Mammalian RNAi Tools

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    ...degrades the mRNA. RISC includes argonaute protein 2 (Ago2), which facilitates cleavage of the target. ... International Journal of Biomedical Science, 13 (2), 48–57. PubMed (Link opens in a new window) . Moore... in therapeutic settings. As a tool in mammalian cell systems, silencing is achieved through the delivery...). Origins and mechanisms of miRNAs and siRNAs . Cell, 136 (4), 642–655. PubMed (Link opens in a new window...
  5. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

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    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(...
  6. Adenovirus Plasmids

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    ...recombining shuttle plasmids Vogelstein 16401 pAdEasy-2 Adenoviral For recombining shuttle plasmids; for use...transgene under CMV promoter Vogelstein 16407 pAdEasy 2-GFP beta-gal Shuttle Test plasmid that contains β-...target a broad range of dividing and non-dividing cell types with almost 100% efficiency. Unlike lentiviruses...recombination of adenovirus plasmids in either mammalian cells or microorganisms, including bacteria and yeast....can then be transfected into mammalian packaging cell lines to generate adenovirus particles. Read our...
  7. AAV Molecular Tools

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    ...diphtheria toxin receptor fused to GFP for studying cell ablation. 2 Jessell , Azim 45580 pAAV-flex-taCasp3-TEVp...constitutive Expression of the tet-off transactivator (tTA) 2 Gradinaru 99120 pAAV-ihSyn1-tTA Inducible Synapsin...mapping 5 Kim Tools for Cell Ablation These AAV encode tools that can be used for cell ablation. ID Name Expression...Transactivators Affinity Purification Neurophysiology Cell Ablation Genome Engineering Labeling Overexpression...expression of designer pro-taCasp3 and TEVp for studying cell ablation. 1, 5 Shah , Wells Genome Engineering Tools...Tools These AAV encode tools that can be used for cell or protein labeling. ID Name Expression System Activity...
  8. Adeno Associated Virus (AAV) for Cell and Gene Therapy

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    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 ...
  9. Cloning Mammalian Cells with the Agarose Method

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    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...
  10. Antibody Plasmid Collection

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    ...phage display. J Immunol Methods. 2000 Aug 28;242(1-2):159-81. Carlos Barbas Vector system for expression...+J phagemid libraries. Immunol Lett. 2015 Oct;167(2):95-102. Joanna Bereta Sybody Generation Toolbox for...expressing these in bacteria, mammalian cells, yeast, plants, or insect cells. Benefits include consistency between... research purposes include animals or hybridoma cells, but are difficult to generate due to expense and...monoclonal antibodies (R-mAbs) expressed in mammalian cells. Monoclonal antibodies are derived from an individual...transiently transfected into mammalian tissue culture cells, the R-mAbs are secreted into the culture media ...
  11. Production of Virus in Insect Versus Mammalian Cells

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

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    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...
  13. Adeno-associated virus (AAV) Plasmids

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    ...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 104963 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...
  14. Zinc Finger Consortium: Zinc Finger Arrays

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    ...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...
  15. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...Seed individual cells in a 96-well plate Day 2–14: Monitor cells for growth and expand cells Day 14–30: Analyze...colony size. Figure 2: Cas9 expression in monoclonal cell lines generated from A549 cells transduced with ... CO 2 incubator Pipet controller Hazardous waste container Microscope Hemocytometer or other cell counter... is about 2 × 10 6 cells in a 10 cm dish. Each 10 cm dish should be seeded in 10 mL DMEM complete, which...transgene expression ( Figure 2 ). Sample Data Figure 1: Generation of monoclonal cell lines from expansion of...Biological Safety Cabinet 0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel...of a 5 cell/mL solution. Calculate the total cells needed: Total cells needed: 10 mL × 5 cells/mL = 50...
  16. Virus Protocol - Generating Stable Cell Lines

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    Protocol
    ...surviving cells. Even in the absence of cell death, the cell media should be changed every 2–3 days to...Day 0: Seed and transduce Cells Day 2–3 (am): Remove media, replace with fresh media containing selection...into two 75 cm 2 flasks, etc. Pro-Tip This selection method results in a polyclonal cell population, meaning...Safety Cabinet 0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel... 200–1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container... the cell media. Cell death by some cells in the culture may adversely affect the surviving cells in the...media to make the cell solution in this step. To seed the cells: Prepare a batch of cells as follows: Dilute...
  17. Four Factors that Differentiate the Stem Cell Field

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    Blog Post
    ... Podcast Interview with Stem Cell Researcher Darrell Kotton 2. Stem cell scientists care about patients...explained that it took 50 people nearly 2 years to create the first stem cell line that could be used in clinical...prevent stem cell lines from becoming the new HeLa cells?), genome engineering and stem cells, and the difficulty...opportunity to see Shinya Yamanaka celebrate the tenth anniversary of his landmark Cell paper showing that it was...generate induced pluripotent stem cells (iPSCs) capable of forming nearly all cell types in the body from otherwise... happening. Stem cells scientists have certainly not lost site of that. 3. Stem cell science is hard, ...otherwise terminally differentiated cells simply by expressing four proteins Oct4, Sox2, Klf4, and cMyc...
  18. CRISPR Guide

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    ...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...
  19. Fluorescent Protein Guide: FRET

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    ...referred to as intermolecular or bimolecular FRET) or (2) conformational changes within a protein where the...standard consisting of ECFP and EYFP connected by 2 GGSGGS repeats pET28CLY3 Peptide linker standard consisting...Protein Resources: Empty Backbones Biosensors Subcellular localization Optogenetics Background Förster ... Constructs to target mTurquoise2 to various subcellular compartments pCEP4CyPet-MAMM Cyan Mammalian Expresses...
  20. Plan Your Experiment

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    ...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...
  21. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

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    Blog Post
    ...the specimen.  2. Fluorescence Activated Cell Sorter (FACs) A machine that sorts cells according to whether...photostability of your fluorescent protein when using 2-photon microscopy as this hasn't been well reported...PMID: 18703742. PubMed Central PMCID: PMC2606057. 2. Chtanova, Tatyana, et al. "Dynamics of neutrophil...labeling T cells, B cells, and antigen presenting cells, scientists have been able to decypher cellular dynamics...tracked by cell sorter giving immunologists the ability to count cells and statistically analyze cell movements...fast T cells could jump from one antigen presenting cell to another. Indeed, we now know that T cells are...2016)? How do dendritic cells migrate to lymph nodes to activate T cells (Celli et al 2008, Kitano et al...
  22. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

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    ...NGG-3′ (“N” can be any nucleotide base, followed by 2 guanines). This limits the use of ABEs as DNA sites... in the promoter region of gamma hemoglobin (HGB1/2) leads to continuous expression of fetal hemoglobin...primary human cells  Cell therapy approaches can use base editors to correct mutations in stem cells and return...primary human cell editing, the researchers tested ABE8 in CD34+ hematopoietic stem cells. ABE8 efficiently... In addition to stem cells the authors targeted 6 genes in primary human T cells using 8 selected ABE8s...to edit primary human T cells efficiently makes it a promising tool for T cell therapy development.  Reducing...be introduced into cells as a plasmid or as mRNA constructs. In primary human cells mRNA delivery of ABEs...
  23. Zinc Finger Consortium: Nuclease Expression Vectors

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    ..., 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 ...
  24. Antibody Guide

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    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...
  25. CRISPR Plasmids - Epigenetics

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    ...Cas enzymes? CasPEDIA is an encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme... time and are potentially heritable in dividing cells. CRISPR Activate and Interfere technologies also...
  26. CRISPR Plasmids - Tagging

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    ...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...
  27. AAV Viral Preps

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    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...
  28. Optogenetics Plasmid Collection

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    ...optogenetic tool is the widely used Channelrhodopsin-2 (ChR2) from Chlamydomonas reinhardtii . ChR2 is a ... optical switch tools to measure and manipulate cells and their biomolecular 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...
  29. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

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    Blog Post
    ...PMC5118044. 2. Kim, Y., et al., The Generation of Human Induced Pluripotent Stem Cells from Blood Cells: An Efficient...blood are reprogrammed immediately after collection [2]. PBMCs display unique epigenetic signatures that ...human-induced pluripotent stem cells from cord blood and peripheral blood cells. Stem Cells, 2013. 31(3): p. 458...possible to reprogram differentiated cells into induced pluripotent stem cells (iPSCs) by the addition of reprogramming... somatic cells that are otherwise difficult to obtain. These can then be used to perform cell therapy ...in vitro. Find stem cell plasmids at Addgene Many sources of adult somatic cells have been probed for...Using Serial Plating of Reprogrammed Cells by Centrifugation. Stem Cells Int, 2016. 2016: p. 1329459. PubMed...
  30. Stem Cell Models for Disease & Open Science: Interview with Darrell Kotton

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    ...develop therapies to treat disease in the lung. Chapter 2: Balancing research with medical practice as an MD...engineered cells in therapy. The lungs are difficult, but other tissues like the retina have cell-based therapies...Chapter 9: Using stem cells as in-vitro models of disease. How Darrell is using stem cells from patients to...sharing reagents, cell lines, data, and knowledge without exclusivity. Chapter 11: The Stem Cell Bank (CReM ...the lungs, and, among other projects, using stem cells to develop in vitro models for lung disease. Darrell...what drives him as a praticing physician and stem cell researcher. Darrell's ultimate goal - to develop...means to treat disease through tissue repair, stem cells, gene therapy, and tissue engineering. How the multidisciplinary...
  31. CRISPR Plasmids - gRNAs

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    Collection
    ...Cas enzymes? CasPEDIA is an encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme...is highly recommended that you determine if your cell type contains the target sequence before using any...
  32. CRISPR Plasmids - Cascade-Cas3

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    Collection
    ...are largely underutilized compared to their Class 2 counterparts like Cas9. This is attributed in part...bound to DNA, and the break can be repaired by the cell’s endogenous repair mechanisms. Cas3 belongs to the...
  33. X-CHIME: Context Dependent Germline Knockout in Immune Cells

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    Blog Post
    ...), 1668. https://doi.org/10.1038/s41467-019-09656-2 More resources on the Addgene blog CRISPR101: Validating...hematopoietic stem cells with a lentiviral vector containing a gRNA. These transduced stem cells can be implanted...Annotation of genes in immune cells typically involves the creation of germline knockout mice, which ...CRISPR-based systems enable gene knockout in immune cells in a high-throughput manner, but these systems have..., ubiquitous knockout of single genes in immune cells in vivo (LaFleur et al., 2019a and LaFleur et al...many genes have different functions in different cell types, at different times, and in combination with... L-CHIME Knocks out a single gene in defined cell types (lineage-based) pXPR_070 Violet-excited...
  34. Bikard Lab - CRISPR Repression Collection

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    Collection
    ...chromosomal integration of DNA. ACS Synthetic Biology, 2(9), 537–541. DOI: 10.1021/sb400021j Figure 1: Multiplexing... CRISPR plasmids. Each point represents a single cell, and each color represents the population obtained...
  35. Resolute Plasmid Collection

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    Collection
    ... funding from the Innovative Medicines Initiative 2 Joint Undertaking under grant agreement No 777372....quality of research output, RESOLUTE expects to accelerate the pace of research in the field of SLCs to ...
  36. CRISPR References and Information

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    Collection
    ...Link opens in a new window) An encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme...library amplification 1 vector system: lentiCRISPR v2 2 vector system: lentiCas9-Blast and lentiGuide-Puro...interest and correspond to edited and non-edited cell populations. For more information, see the Hsiau...138 KB Musunuru CRISPRs in human pluripotent stem cells pCas9_GFP ; gRNA empty vector Protocol at StemBook...Bauer Protocol for Genomic Deletions in Mammalian Cell Lines pSpCas9(BB) (pX330) PDF, 330 KB Parrott NEBuilder...
  37. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

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    Blog Post
    ...(I), or sparse (S) labeling of the cell population. Figure 2: Dense (c), intermediate (d), and sparse...single genetically-defined subset of cells. By labeling fewer cells in a population, it’s easier to visualize...non-overlapping cells. While transgenic animals are commonly used to specifically manipulate a cell type of interest...expressed. In cells where the stop cassette remains, the effector is not expressed and the cells are not labeled...sparse labeling of the cell population that expresses SPARC and the fraction of cells that will be labeled... interest in an organism, all cells of that type are affected. Previous tools to overcome this restraint... of the gene construct. A predictable number of cells will express the effector or retain the stop cassette...
  38. AAV for Neuronal Tracing

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    ...G-protein is delivered to the starter cell. (2) The starter cell now expresses TVA and G-protein. EnvA-pseudotyped...spread from this cell to synaptically connected cells (meaning, it can spread to cells that are projecting...infect a single "starter" cell, but new virus cannot spread to any neighboring cells. To enable monosynaptic...provided to the starter cell in trans , via delivery by an AAV. Since this starter cell now has all the viral...projecting to the starter cell). Once in the secondarily infected cells that lack the G protein, the virus ...connected cells. To further refine the targeting of G-deleted rabies to specific starter cells, the tropism...is delivered to the starter cell, and specifically targets the starter cell via the EnvA receptor. RABV...
  39. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

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    Blog Post
    ...SARS-CoV-2 spike pseudotyping, visit the SARS-CoV-2 Pseudotyped Virus collection page. SARS-CoV-2 detection...of SARS-CoV-2 entry and become upregulated during viral infection.  Expression of SARS-CoV-2 viral proteins...pseudotyping The SARS-CoV-2 virus depends on its spike protein to bind ACE2, the host cellular receptor. To more...point for SARS-CoV-2 researchers to investigate how the virus interacts with human cells. Addgene’s mammalian...genes are important for SARS-CoV-2 entry into human lung epithelial cells (Biering et al., 2021). In their...Wesemann DR (2021) Memory B cell repertoire for recognition of evolving SARS-CoV-2 spike bioRxiv. https://...labs in over 75 countries. Figure 1: SARS-CoV-2 proteins and other components. Image from Maya Kostman...
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