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We narrowed to 819 results for: cell

Showing: 571 - 600 of 819 results

  1. What's New in CRISPR - September 2019

    Type
    Blog Post
    ...small Cas protein, is active in E. coli and human cells Two years ago, Jennifer Doudna’s lab discovered ...demonstrating that CasX is active in both E. coli and human cells. Using the structure of CasX at eight different ...

  3. Plasmids 101: Degron Tags

    Type
    Blog Post
    ...tag and small molecule. It does work in mammalian cells, mice, and even yeast, though (we have a kit!). ...and complete degradation of proteins in mammalian cells. Proc Natl Acad Sci U S A. 2012 Dec 4;109(49):E3350...Tags Plasmids for Endogenous Gene Tagging in Human Cells Antibodies 101: Epitope Tags  ...uses a small molecule, a tag, and the endogenous cellular degradation machinery to function. The dTAG is...

  4. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ... check viral transgene expression in transduced cells whenever possible. This post will describe our workflow...fragments of DNA from the helper plasmids or the cell genome can be packaged inside the vectors during...earlier, it is common to find DNA from the packaging cell genome, bacterial genomes, cloning vectors, and ...

  5. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ... localization of that biomolecule inside living cells. FPs classified by the emission color (emission ...proteins and their applications in imaging living cells and tissues”  Physiol. Rev. 90:1103. Wu et. al. ... nuclear localization, and dynamics” Curr. Opin. Cell. Biol. 23:310. .... Therefore, these can be used as “timers” for cellular processes following their activation. The four...Biosensors: Throwing color lights on molecular and cellular processes” Curr. Protein. Pept. Sci. 9(4):338....

  7. New CRISPR Web Resources and #12DaysofCRISPR Recap

    Type
    Blog Post
    ...predict that we’d be able to edit RNA in mammalian cells with CRISPR? I certainly didn’t, but now we can ...Improve HDR Efficiency for CRISPR Editing in Human Cells Day 3: Cpf1: Cpf1: A New Tool for CRISPR Genome ...

  8. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ... . Steps of an IP: 1) Protein extracts from cells or tissues contain a complex mix of proteins seen...common immobilization methods utilizes bacterial cell wall proteins, Protein A and Protein G, which bind...does not. Common negative controls include knockout cell lines or tissues that do not express the protein...

  9. Analyzing CRISPR Editing Results with ICE from Synthego

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    Blog Post
    ...score (KO-score) which represents the proportion of cells that have either a frameshift or 21+ bp indel. This...Following delivery of CRISPR components into target cells, genomic DNA from both edited and unedited (control... score. KO Score - Represents the proportion of cells that have either a frameshift or 21+ bp indel. This...

  10. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    ...triple colocalization is possible even when staining cells with three IgG1 antibodies. Antibodies were individually...workflow yields similar localization patterns as cells stained for one target (compare nanobodies staining...–rabbit IgG secondary nanobodies. The Journal of Cell Biology. PubMed PMID: 29263082 Find plasmids from...

  11. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    ...example, "while it is possible to grow liver stem cells and liver-like mini-organs or organoids, neither...combine bioengineered scaffolds with patient stem cells to generate compatible functional livers.” Why are... optimization of fluorescent proteins,  and live cell imaging approaches. Erik is also an avid long distance...

  12. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ...mismatched translation rates and potential effects on cell growth, even hosts producing alternative tRNAs may...inclusion bodies that will generally segregate with the cell pellet during purification procedures. Even if you...a means to fine-tune gene expression." Molecular cell 59.2 (2015): 149-161. PubMed PMID: 26186290. PubMed...reasons include: Metabolic pressures - it takes cellular resources to produce tRNAs that recognize different...

  13. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ...Control of Gene Expression. Cell 152:1173–1183 . https://doi.org/10.1016/j.cell.2013.02.022 Ronda C, Pedersen... is primarily a means of selection that targets cells in which homologous recombination has not occurred...ensures high editing efficiency; the only non-edited cells to survive have inactivating mutations in the Cas9...one or more genes in E. coli. and in a mammalian cell line (Qi et al., 2013). Two important features of...the bacteria are unable to resolve them, and the cells die.  This could be applied to diseases where the...

  14. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ...activator domain (VP64 or p65). Upon expression in the cell, the dCas9-CIB1 fusion binds to the target DNA sequence...activator domain VP64. Using this system in HEK293T cells to induce expression of human IL1RN, the researchers...protein with three red fluorescent tags in U2OS cells. Upon light activation, dCas9 is released within... silencing of transferrin receptor CD71 in HeLa cells.   Comparison of photoactivatable CRISPR strategies...Spatiotemporal Control of CRISPR/Cas9 Function in Cells and Zebrafish using Light‐Activated Guide RNA. Angew...encoded tools, such as microbial opsins, to control cellular activities using light. In 2015, scientists combined...

  15. Brain Initiative Collection

    Type
    Collection
    ...real-time interrogation and monitoring of brain cells and circuits during behavior. Long desired by researchers...A somatic channelrhodopsin, which enables single cell, single millisecond resolution optogenetics. Human...A somatic channelrhodopsin, which enables single cell, single millisecond resolution optogenetics. Cre-dependent...genetically encoded dopamine sensor RdLight1 in mammalian cells. Lin Tian 125708-AAV9 pAAV-hSynapsin1-RdLight1 Expresses...neurons Yulong Li 158757-AAV8 pAAV-CAG-SomaGCaMP6f2 Cell body-targeted GCaMP6f, under CAG promoter: GCaMP6f... Edward Boyden 158757-AAV9 pAAV-CAG-SomaGCaMP6f2 Cell body-targeted GCaMP6f, under CAG promoter: GCaMP6f...

  16. Genetic Code Expansion

    Type
    Collection
    ...expanding the genetic code in bacteria and mammalian cells....codons, and tRNA synthetases are introduced into the cell on plasmids and the new amino acid is introduced...Once these plasmids have been introduced in the cells, the non-canonical amino acid can be incorporated...match the growth medium, ncAA concentration, and the cell lines used previously. Remember that the orthogonal... – GFP for E. coli or mCherry-GFP for mammalian cells. You should also express the protein with and without...tRNA synthetase for use in E.coli and Mammalian Cells. Many of the plasmids also contain one or more copies...

  18. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    ... a common thread at the meeting. The new Somatic Cell Genome Editing program, run through the NIH Common...Improve HDR Efficiency for CRISPR Editing in Human Cells Hands-on tips for repair template design from guest...to learn much more about cellular DNA repair pathways. An emphasis on cellular repair post-CRISPR cleavage..., which alter genomic DNA without the need for cellular HDR.   Having had time to digest my notes, since...

  20. Using Addgene's New Viral Service for Your Research

    Type
    Blog Post
    ... can be used to deliver fluorescent proteins to cells as determined by the serotype. You can use these...determine if your AAVs are getting to the right cells using your particular delivery technique and conditions...

  21. FastCloning: A Newer, Simpler Cloning Technique

    Type
    Blog Post
    ...properly. Directly transform chemically competent cells with the digested mixture. There is no need for ...purification before transformation. After transforming cells, you should perform colony PCR with vector specific...

  23. CRISPR References and Information

    Type
    Collection
    ...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...

  24. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ...experiments possible in a wide range of organisms and cell lines. In C. elegans, one of the model organisms... efficiency. Finally, as shown in Drosophila S2 cells (Böttcher et al., 2014), inactivation of non-homologous... community. Developments in mammalian and yeast cells using modified Cas9 proteins to regulate gene expression...homologous recombination donors in cultured Drosophila cells. Nucleic Acids Research 42:e89–e89 . https://doi.org...

  25. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    ...delivered as a pooled library via a lentiviral vector to cell lines that either stably expressed Cas9 or that ...almost all genes involved in the production of a cell surface glycan that’s required for ricin uptake....reveals pathways underlying ricin susceptibility." Cell 152.4 (2013): 909-922. Pubmed PMID: 23394947. Pubmed...

  26. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    ...optogenetics, ICSS can now be done with neuronal cell-type and temporal precision. In ICSS, a test chamber...differential participation in reward circuitry." Cell reports 8.6 (2014): 1857-1869. Pubmed PMID: 25242321...Dynamics of retrieval strategies for remote memories. Cell. 2011;147(3):678-89. Pubmed PMID: 22179551. Pubmed... remote contextual memories. This study is an excellent example in which the authors started with a brain... research. Derek P. Simon received his PhD in Cellular and Molecular Biology from the University of Michigan...

  27. Fluorescent Protein Guide: Activity Regulation

    Type
    Collection
    ...components or take advantage of existing GFP-lines for cell-specific manipulation. Plasmids Fluorescent Protein...Fluorescent Protein Resources: Empty Backbones FRET Subcellular localization Biosensors Optogenetics Background...

  29. Cultivating Community Science at BosLab

    Type
    Blog Post
    ...engineering project at BosLab. Once he optimizes plant cell culture and transgenesis he wants to share his knowledge... in an Open Source Lab: from CRISPR to Cats Stem Cell Models for Disease & Open Science: Interview with...

  30. Antibodies 101: Epitope Tags

    Type
    Blog Post
    .... This tag is not suitable for use in apoptotic cells as Caspase3/7 both cleave after the DVPD sequence...fluorescent marker. The GFP tag is ideal for live cell imaging, FRET, flow cytometry, or immunofluorescence...
Showing: 571 - 600 of 819 results