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  1. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...simpler to implement in genome-wide screens over cDNA libraries. However, using CRISPRa to target individual...highlight a key advantage of CRISPRa over traditional cDNA overexpression: isoform-specific gene expression...— at least, more straightforward than generating cDNA for each gene isoform individually. To highlight...amenable to FACS.  Thus, the researchers inserted human CD8, a gene that encodes a surface protein for which...application can be found here.  Why CRISPRa? Although cDNA overexpression approaches may still be preferable...strategies) and then consider whether traditional cDNA overexpression is suited to your experimental goals...
  2. Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection

    Type
    Blog Post
    ...large scale, broadly available collection of mutant cDNA clones corresponding to cancer variants found in... genome sequencing studies (as well as reference cDNA clones mapping to well-annotated cellular pathways...Addgene. These include 634 sequence confirmed mutant cDNA clones corresponding to cancer variants derived ...paper Systematic morphological profiling of human gene and allele function via Cell Painting. Rohban...
  3. CRISPR Activation: A Practical Guide

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    Blog Post
    ...they want to activate, we would recommend using a cDNA overexpression vector rather than going through ...genetic screening. gRNAs targeting every gene in the human genome for example, can be made easily and cheaply...protein structure and function (Pan et al., 2018). cDNA libraries, which consist of plasmids that over-express...and deliver when compared to gRNAs. Additionaly, cDNAs cannot be used to study in cis regulation and also...
  4. Lentivirus Plasmids

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    Collection
    ...expression of cDNA. Puro selection. Shih 25737 pSLIK-Hygro 3rd Tet-based inducible shRNA or cDNA expression...RD114 Env GP and MLV 4070A Env GP. Good for work in human MSCs. Use packaging construct encoding Tat. Reiser...EGFP 3rd CMV-driven EGFP fusion; can be used for cDNA expression; puro resistance Sabatini 14883 FUGW ...FUGW 3rd hUbC-driven EGFP; can be used for cDNA expression Baltimore 12247 pLVTHM 2nd EF-1a-driven GFP and...plasmid with constitutive or inducible expression of cDNAs, shRNAs or miRNAs, and use a wide variety of drug...3rd Tet inducible gateway destination plasmid for cDNA expression Ramalho-Santos 24129 pULTRA 3rd bi-cistronic...3rd Gateway plasmid for CMV driven expression of cDNA. See article for alternative versions of this plasmid...
  5. Viral Vectors 101: Viral Vector Elements

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    Blog Post
    ...expression construct, to cells/organisms. This is often cDNA, shRNA, or something similar, and is contained on...infrequent genome integration at the AAVS1 locus in humans. Cap is a structural capsid protein that helps ...-293) during production. Fun fact: There are 57 human adenovirus types, but most vectors are based on ...
  6. PITChing MMEJ as an Alternative Route for Gene Editing

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    Blog Post
    ...harbor locus” of the human genome, as shown by Dalvai et al., who used HDR to insert cDNA constructs into ... Approach for Mapping Multiprotein Complexes in Human Cells.” Cell Rep. 13: 621-633. PMID: 26456817. ...
  7. Cancer Research Plasmids and Resources

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    Collection
    ...cell. p53 p53, the most frequently mutated gene in human cancer, is a transcription factor and tumor suppressor...vectors from the Sabatini & Wood Labs containing cDNAs for studying oncogenic signaling pathways. ClonTracer...
  8. MXS Chaining

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    Blog Post
    ... 3 mCherry 587nm/ 610 nm human β-Actin Actin 4 Citrine 516nm/ 529nm human α-Tubulin Tubulin Repeatable...of these species and can often be used to clone cDNAs from them. These enzymes might not, however, be ...
  9. Plasmids 101: Gateway Cloning

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    Blog Post
    ...desired constructs: lentiviral expression of the human KRAS gene in mammalian cells. STEP 1: Generate an...different ways to generate our desired entry clone - human KRAS flanked by attL sites.  Method A: recombination...types of DNA fragments may be cloned: PCR fragments, cDNA or Genomic DNA and is available for all kind of ...
  10. Plasmids 101: Using Transposons in the Lab

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    Blog Post
    ...genome. In humans, if a transposon jumps into a gene, it could be mutagenic and result in human disease....prokaryotes and eukaryotes, and make up ~45% of the human genome (Munoz-Lopez et al., 2010).  So why do transposons.... SB is active in vertebrates and integrates in human cells at rates similar to retroviral vectors. The..., plant, insect, and mammalian cells, including human cells. piggyBac is biased to integrate at transcription...Beauty, piggyBac has an integration efficiency in human cells similar to retroviral vectors. The hyperactive...vertebrates and has lower efficiency of integration in human cells than piggyBac and Sleeping Beauty. Minimal...for lentivirus). Large cargos, such as the ~11 kB cDNA for dystrophin, the gene mutated in muscular dystrophy...
  11. CRISPR Plasmids - Tagging

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    Collection
    ... pFETCh_Donor CEBPB Human FLAG pFETCh_CEBPB PX458_CEBPB_1 PX458_CEBPB_2 AHR Human FLAG pFETCh_AHR PX458...AHR_1 PX458_AHR_2 ARID4B Human FLAG pFETCh_ARID4B PX458_ARID4B_1 BCL6_iso1 Human FLAG pFETCh_BCL6_iso1 PX458..._BCL6_iso1_2 CEBPG Human FLAG pFETCh_CEBPG PX458_CEBPG_1 PX458_CEBPG_2 RAD21 Human FLAG pFETCh_RAD21 PX458...PX458_RAD21 GABPA Human FLAG pFETCh_GABPA PX458_GABPA_1 PX458_GABPA_2 GABPB1_iso2 Human FLAG pFETCh_GABPB1...iso2 PX458_GABPB1_iso2_1 ATF1 Human FLAG pFETCh_ATF1 PX458_ATF1 CEBPA Human FLAG pFETCh_CEBPA PX458_CEBPA...PX458_CEBPA_2 CREB1 Human FLAG pFETCh_CREB1 PX458_CREB_1 PX458_CREB_2 TGIF2 Human FLAG pFETCh_TGIF2 PX458...PX458_TGIF2_2 SSRP1 Human FLAG pFETCh_SSRP1 PX458_SSRP1_1 PX458_SSRP1_2 ZNF146 Human FLAG pFETCh_ZNF146...
  12. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

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    Blog Post
    ...easily isolated from donors. Apart from fibroblasts, human keratinocytes from hair pluck, peripheral blood ...types have been successfully differentiated from human iPSCs. Below is a list of some of these cell types... Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell, 2007. 131(...Induced pluripotent stem cell lines derived from human somatic cells. Science, 2007. 318(5858): p. 1917... et al., Efficient generation of transgene-free human induced pluripotent stem cells (iPSCs) by temperature-sensitive...PubMed Central PMCID: PMC3161531. 7. Yu, J., et al., Human induced pluripotent stem cells free of vector and...to pluripotency and directed differentiation of human cells with synthetic modified mRNA. Cell Stem Cell...
  13. Fluorescent Proteins 101: When GFP lets you down

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    Blog Post
    ...proteins are made by connecting its DNA code to the cDNA encoding for another protein. Effectively, it equips... al. "Dynamic imaging of genomic loci in living human cells by an optimized CRISPR/Cas system." Cell 155.7...
  14. Delivery Methods for Generating iPSCs

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    Blog Post
    ... They contain only a eukaryotic promoter and the cDNA(s) to be expressed and they don’t integrate or replicate... of these functional tests aren’t possible with human iPSCs, the the ability to generate teratomas is ...Robert Lanza, and Kwang-Soo Kim. "Generation of Human Induced Pluripotent Stem Cells by Direct Delivery...and Mahendra S. Rao. "A Review of the Methods for Human iPSC Derivation." Methods in Molecular Biology Pluripotent... Wu. "A nonviral minicircle vector for deriving human iPS cells." Nature Methods 7, no. 3 (2010): 197-...reprogramming factors: Mouse Expression Vectors Human Expression Vectors Carey et al Hockemeyer et al...
  15. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ...constitutive (CMV, SV40, SFFV) or inducible (CMV-TO) cDNA expression, sgRNA (U6) expression, shRNA/shRNA-miR30...CreERT2 gene deletion, (3) simultaneous expression of cDNAs and shRNAs with a reporter gene, or (4) multiple...thermophilus which have been used for gene editing in human cells without cross-talk in cognate sgRNA binding...understanding the spatio-temporal regulation of human genes. The plasmids are all available at Addgene...These three enzymes were chosen because they cut human and mouse coding regions infrequently; any MluI,...understanding the role of long noncoding RNAs (lncRNAs) in human health and disease. A primary question in their ...He has also constructed a library of eGFP-tagged human ORF clones to allow testing and comparison of different...
  16. Neurodegeneration Research Collection

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    Collection
    ...astrocytes. Endo et al Science. 2022 Nov 4. Base edit human mitochondrial DNA using mitoBEs . Yi et al. Nat ...(Link opens in a new window) such as huntingtin cDNAs, antibodies, and cell lines. Neurodegeneration Method...
  17. 27 Hot Plasmids from 2016

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    Blog Post
    ... both human and fly versions. Arnold, et al. Science. 2013. PubMed PMID: 23328393 Wu lab human lentiviral...   Human kinase domain constructs for automated bacterial expression The misregulation of human kinases...Lab recently compiled the Human Kinase Domain Constructs Kit, a library of human kinase domain constructs...gRNAs targeting a total of 20,121 genes within the human genome (the identity of the genes and target sequences...repository that can be used to perform screens in human cells. General information regarding how to use ...receptors (GPCRs) yet one third of the GPCRs in the human genome are orphan receptors meaning their ligands...allows researchers to interrogate the druggable human GPCR-ome using a quick, affordable, and ubiquitous...
Showing: 1 - 20 of 29 results