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We narrowed to 72 results for: h cas9 protein

Showing: 61 - 72 of 72 results
  1. Plasmids 101: Screens vs. Selections

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    Blog Post
    ...players in a biological process. Players can be proteins, genes, enzymes, drugs, or anything else that ... Genetic screens in human cells using the CRISPR/Cas9 system. Science, 343(6166): 80-84 (2014). doi: https...https://doi.org/10.1126/science.1246981. Leemhuis, H., Kelly, R.M., Dijkhuizen, L. Directed evolution of...
  2. Lentiviral Vector Uses and Overview

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    Blog Post
    ...that couldn’t package itself due to a few missing proteins. The envelope plasmid contained a viral envelope...for viral production: Gag (structural precursor protein), Pol (polymerase), Tat (viral transactivator for...export of transcripts). A different viral envelope protein (Env), usually from VSV-G due to wide infectivity...Screening Read about Genome-Wide Screening Using CRISPR/Cas9 Additional Resources on Addgene.org Check out ...of a self-inactivating lentivirus vector. Miyoshi H, Blömer U, Takahashi M, Gage FH, Verma IM. J Virol...
  3. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

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    Blog Post
    ...recognition sequences of an exogenous DNA-binding protein such as LexA into the target locus, which is the... DNA-binding molecule technologies such as TAL proteins and CRISPR became available, I was very excited...easily isolated for identification of associated proteins, RNAs, and other genomic regions. I believe that...analyze signal-induced nuclear translocation of proteins in 2004. In addition, we have recently developed...Addgene Alerts box. References Identification of Proteins Associated with an IFNgamma-Responsive Promoter...genomic regions and identification of associated proteins by engineered DNA-binding molecule-mediated chromatin...Expression System for enChIP Using CRISPR. Fujita T, Fujii H. PLoS One. 2014 Jul 22;9(7):e103084. doi: 10.1371/...
  4. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

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    Blog Post
    ...test varies in the sequence of steps and the Cas protein used. Here, all the components needed for the reaction...coronavirus using CRISPR-based SHERLOCK (Specific ​H​igh ​s​ensitivity ​E​nzymatic ​R​eporter un​LOCK​ing...    References Azhar, M., Phutela, R., Ansari, A. H., Sinha, D., Sharma, N., Kumar, M., … Maiti, S. (2020... Gopez A, Hsu E, Gu W, Miller S, Pan C-Y, Guevara H, Wadford DA, Chen JS, Chiu CY (2020) CRISPR–Cas12-...Approach: FnCas9 Editor Linked Uniform Detection Assay (FELUDA) is a technique that uses FnCas9, a protein...sensitivity of FnCas9, this is also a good way to diagnose single nucleotide variations. Also, FnCas9 cleaves...field-deployable nucleobase detection and identification using FnCas9 (FELUDA) Sensitivity: – 110 femtomolar  Time: Not...
  5. Adeno-associated Viruses (AAVs) for Genome Editing

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    Blog Post
    ... hampered by the fact that even small Cas9 homologs like SaCas9 and their requisite gRNAs take up essentially...: PMC3277219. 6. Ohmori, Tsukasa, et al. "CRISPR/Cas9-mediated genome editing via postnatal administration...PMCID: PMC5482879. 7. Bak, Rasmus O., and Matthew H. Porteus. "CRISPR-mediated integration of large gene...et al 2017 use a dual vector system composed of SaCas9 and a gRNA in one AAV and a repair template in ...devised methods to deliver CRISPR components as ribonucleoproteins (RNPs) that can be supplied separately from...
  6. CRISPR Guide

    Type
    Guide
    ...inhibiting Cas9’s endonuclease domain, or inducing degradation of the Cas9 protein. Most Acr proteins are specific...Gelsinger, D. R., Vo, P. L. H., Klompe, S. E., Ronda, C., Wang, H. H., & Sternberg, S. H. (2024). Bacterial genome...Vo, P. L. H., Ronda, C., Klompe, S. E., Chen, E. E., Acree, C., Wang, H. H., & Sternberg, S. H. (2020)....constructs with Cas9 fused to proteins that modulate HDR or NHEJ components, like miCas9 , Cas9-DN1S , or others..., such as Cas9 or Cas12a. These are identified in their names — AcrII proteins inhibit Cas9 (CRISPR Type... available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements...Z., Xie, H., Liu, D., Mo, H., Zhou, R., Lang, B., Meng, B., Weng, H., & Hu, Z. (2022). FrCas9 is a CRISPR...
  7. RNA Interference in Plant Biology: New Tools for an Old Favorite

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    Blog Post
    ...in a direct or unbiased way at the DNA, RNA, or protein level. The “correct” choice of tool requires careful...unmatched—unlike CRISPR experiments that require exogenous protein, successful induction of RNAi only requires the...scored. However, determining the transcript or protein abundance of your specific target required physical...Bush J, McCormack M, Sheen J (2013) Comprehensive Protein-Based Artificial MicroRNA Screens for Effective...Bassik MC (2016) Systematic comparison of CRISPR/Cas9 and RNAi screens for essential genes. Nat Biotechnol...Fälth Savitski M, Grandi P, Jakob P, Michon A-M, Sun H, Tessmer K, Bürckstümmer T, Bantscheff M, Steinmetz...
  8. Plan Your Experiment

    Type
    Guide
    ...copying from a DNA donor template Cas9 or Cas9 fusions with proteins that promote HDR Requires a repair...truncated protein. Targeting an exon coding for an essential protein domain could abolish protein activity...breaks Base editor (dCas9 or Cas9 nickase fusions with different base editing proteins) Many base editors...). You will decide how to express Cas9, the delivery system for Cas9 and the guide RNA (gRNA), the design...non-homologous end joining (NHEJ) at the cut site Cas9 or Cas9 nickase High-fidelity Cas enzymes increase specificity...site. Consider alternative Cas9 enzymes with different PAM sequences or Cas9 enzymes with flexible PAM ...promoter region of the gene of interest, where the Cas9 protein can block transcription from ever initiating...
  9. Hot Plasmids: Summer 2024

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    Blog Post
    ... DNA-targeting proteins, so the team created versions using dCas9, zinc finger proteins, and transcription... of proteins captured on magnetic beads (Arimura et al., 2024). In MagIC-cryo-EM, target proteins (expressed...adding Late Embryogenesis Abundant (LEA) proteins, small proteins that naturally adsorb to the AWI, can ... “miniCRISPR” version, are much smaller proteins than SpyCas9, allowing them to be packaged in AAV vectors...with a histone tail (H3K4me0) to a DNA-binding protein (dCas9 shown here). D3L recruits the endogenous, autoinhibited...for enrichment and structure determination LEA proteins protect samples during cryo-EM grid freezing Engineered...biotinylated SpyTag-SpyCatcher linkers and spacer proteins ending with a GFP nanobody (Figure 1). This enrichment...
  10. Adenovirus Guide

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    Guide
    ..., N., Song, W. X., Chen, J., Sharff, K. A., Luu, H. H., Haydon, R. C., Kinzler, K. W., Vogelstein, B.,... the viral particle and are only composed of a protein capsid and the viral genetic material inside. Adenoviral...after the virus enters the host cell, and encode proteins necessary for viral DNA replication and host cell...replication starts and primarily encode for structural proteins needed to assemble icosahedral capsids and build...viral vectors in combination with foreign capsid proteins. Researchers have further refined the tropism ... or when needing to package larger cargo. CRISPR-Cas9 Delivery As compared to other viral vectors, rAdV...provide a large transgene packaging capacity. Large protein-coding genes and complex gene expression cassettes...
  11. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...region encodes three structural capsid proteins or viral proteins (VP1, VP2 and VP3) that form the icosahedral...Addgene. CRISPR/Cas9 Delivery rAAV vectors are a popular choice for the delivery of CRISPR/Cas9 elements, making... split inteins, proteins that carry out a naturally occurring process known as protein trans-splicing.... genome editing. Packaging the Cas9 from Streptococcus pyogenes (SpCas9) and a gRNA together into an rAAV...this obstacle, has been to use smaller Cas9 orthologs, such as SaCas9 from Staphylococcus aureus . The Feng... the viral particle and are only composed of a protein capsid and the viral genetic material inside. The...region encodes four non-structural replication proteins (Rep78, Rep68, Rep52, and Rep40), which are involved...
  12. Lentiviral Vector Guide

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    Guide
    ...the Rev protein binds. Psi (Ѱ) in cis RNA packaging signal; recognized by nucleocapsid proteins and essential...j.omtm.2021.03.018 PMID: 33981779 Kim, H. S., Hwang, G., Lee, H. K., Bae, T., Park, S., Kim, Y. J., Lee...PMID: 33768124 Marshall, H. M., Ronen, K., Berry, C., Llano, M., Sutherland, H., Saenz, D., Bickmore, W... vesicular stomatitis virus G glycoprotein (VSV-G), an envelope protein with broad tropism used to pseudotype...infectivity) VSV-G (vesicular stomatitis virus G protein) is the most common envelope gene in all systems...third-generation systems do not require the HIV protein Tat to activate the promoter on the transfer plasmid..., J. G., & Zhang, F. (2013). Genome-Scale CRISPR-CAS9 knockout screening in human cells. Science , 343...
Showing: 61 - 72 of 72 results