Sam
9 months ago
Method

Materials and Methods

Cloning, expression and purification The S. pyogenes cas1 and cas9 genes were cloned into pET28a vectors with a C-terminal (His)6 tag (Supplementary Table S1). The S. pyogenes cas2 and csn2 genes were cloned into pET28a and pHMGWA vectors, respectively, each of which contain an N-terminal (His)6-maltose binding protein (MB...
Sam
11 months ago
Method

Methods

Cell lines Human 293T/17 (ATCC No.: CRL-11268) cells were cultured in DMEM (Sigma-Aldrich) supplemented with 10% fetal calf serum (FCS; Sigma-Aldrich) and 2 mM L-glutamine (Sigma-Aldrich) at 37°C and 5% CO2. Human BlaER1 cells (a kind gift of Thomas Graf) (29) cells were grown in RPMI (Sigma-Aldrich) supplemented with 10% ...
Sam
11 months ago
Method

Methods

Strains, plasmids, phage, and oligonucleotides Bacterial strains, plasmids, and phage used in this study, including descriptions and sources, are provided in Supplementary Table 1. Oligonucleotides used in this study are provided in Supplementary Table 2. Minimum inhibitory concentration (MIC) assay Cells were prepared b...
Sam
11 months ago
Method

Methods

T cell editing Whole blood was obtained with donor consent from the Israeli Blood Bank (Magen David Adom, Sheiba Medical Center) in accordance with Tel Aviv University Review Board. PBMCs were extracted using Lymphocyte Separation Medium (mpbio) and cryopreserved until subsequent use. Following thawing, cells were activate...
Sam
11 months ago
Method

Methods

Cells and viruses BSC-40 cells were obtained from American Type Culture Collection (ATCC, CRL-2761) and were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Gibco 11965-092) supplemented with 10% (vol/vol) fetal bovine serum (FBS, Atlanta biologicals S11150H), 1% (vol/vol) glutamine (Gibco 35050-061), and 1% (vol/vol) ...
Sam
11 months ago
Method

Methods

Plant materials Seeds of Nicotiana tabacum cv. ‘SR1’ were obtained from the Leaf Tobacco Research Center. Nicotiana benthamiana, N. tabacum ‘SR1’, and Torenia fournieri cv. ‘Blue and White’ plants were grown in soil in a green room at 25–30°C under long-day conditions (16 h light/8 h dark), and Solanum lycopersicum cv. ‘Mi...
Sam
11 months ago
Method

Methods

Cell Lines and Antibodies U937 and HEK293 cell lines were obtained from American Type Culture Collection (ATCC). U937 cells were cultured in RPMI 1640 with HEPES (Gibco 22400-089) supplemented with 10% fetal bovine serum, defined (HyClone, GE Healthcare SH30070.03); 50 U/mL penicillin, 50 μg/mL streptomycin (Gibco 15070-06...
Sam
11 months ago
Method

Methods

Reagents All chemicals were purchased from Sigma-Aldrich (St. Louis). All enzymes were purchased from New England Biolabs (Beverly, MA). All synthetic DNAs were purchased from Integrated DNA Technologies. C. elegans strains C. elegans strains were cultured using standard methods (Brenner, 1974) on nematode growth media (...
Sam
11 months ago
Method

Methods

Animal husbandry and ethics Medaka (Oryzias latipes) (Iwamatsu, 2004, Naruse et al., 2004, Kasahara et al., 2007) were maintained as closed stocks in a fish facility built according to the European Union animal welfare standards and all animal experiments were performed in accordance with European Union animal welfare guid...
CRISPR/CAS9 endogenous protein Molecular Biology
Find your community. Ask questions. Science is better when we troubleshoot together.
Find your community. Ask questions. Science is better when we troubleshoot together.
Community Guidelines

Sci Find is a place for experimentation beyond the publication. We are a growing collaborative community of scientists and experimentalists from around the world who believe science is better when we work together.

  • Help us keep Sci Find a safe space for productive and collaborative conversations. Please be kind and respect your fellow community members. If you see abusive or alarming content, please report it to admin@scifind.net, and our team will take appropriate actions.
  • All information posted to this page is public and Open Access. Learn more about our commitment to Open Access by visiting our Docs.
  • We encourage our community to have collaborative troubleshooting discussions, and to share tips and tricks, experimental methods, resources, and opportunities.
  • You must be a member to create or engage with content. Sign Up to create your profile and join the community for free.
  • Follow a guild and create a post. Learn more about guilds here.
  • Post titles should be descriptive and include descriptive tags. Check out our tagging guidelines here.

Have a question?

Contact support@scifind.net or check out our support page.