The Cas9 Inducible (Tet-On) iPS Cell Pool expresses the Cas9 (Streptococcus pyogenes CRISPR associated protein 9) gene under the control of a tight TRE tetracycline-inducible promoter, introduced into PBMC-derived human iPS cells via lentiviral transduction [Cas9 Lentivirus (inducible TET on), BPS Bioscience #78794]. Cas9 expression can be induced with doxycycline treatment, allowing temporal control of its expression. Cas9 Inducible (Tet-On) iPS cells can be transduced with single-guide RNA (sgRNA) targeting gene(s) of interest to generate genetically engineered cell pools or cell lines.
Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.
●
Synonyms
iPSC
●
Product Data Gallery
Oct4 Expression in Cas9 Inducible (Tet-On) iPS Cell Pool by flow cytometry.
Cas9 protein expression analysis by Western Blot in Cas9 Expressing iPS Cell Pool.
The cell pool has been screened to confirm the absence of Mycoplasma species.
Background
Cas9 (Streptococcus pyogenes CRISPR associated protein 9) is an endonuclease enzyme that, when recruited to a specific DNA sequence by the appropriate sgRNA (single guide RNA), introduces a double stranded break into the DNA. This double stranded break can then be repaired through either Non-Homologous End Joining (NHEJ) or Homologous Recombination (HR). While NHEJ is an error prone process and causes insertions or deletions which may result in functional inactivation of the target gene, HR, in conjunction with a single stranded ssDNA repair construct, can be used to introduce mutations at specific base pair(s). Gene modifications introduced via Cas9 are now used in multiple fields of research, aimed at both understanding cellular mechanisms and developing therapeutic solutions.
The discovery by Yamanaka and colleagues in 2007 that 4 factors were sufficient to reprogram terminally differentiated fibroblasts into pluripotent stem cells launched the advent of human induced pluripotent stem (iPS) cell technology. These human iPS cells are capable of both self-renewal and differentiation down all three germline lineages and provide both a tool to model human development and disease in the relevant differentiated human cell types. Together with CRISPR/Cas9 technology, gene edited iPS cells provide a unique opportunity to study disease-specific mutations or knockout genes of interest, alongside isogenic control cell pools or cell lines.
Storage/Stability
Cells are shipped in dry ice and should immediately be thawed or stored in liquid nitrogen upon receipt. Do not use a -80°C freezer for long term storage.
As this is a cell pool and not a cell line, BPS Bioscience cannot guarantee the stability of the genetic modifications over time. Clonal selection can be performed. We recommend freezing cell vials very early on and growing the cells for a limited number of passages. Cells should be cultured using Growth Media, which contain selection antibiotics.
Growth Media
For best results, the use of validated and optimized media from BPS Bioscience is highly recommended. Other preparations or formulations of media may result in suboptimal performance.
Media Required for Cell Culture
iPSC Thaw Medium: mTeSR™ Plus supplemented with 1% Penicillin/Streptomycin.
Complete iPSC Thaw Medium: mTeSR™ Plus supplemented with 1% Penicillin/Streptomycin and 1 µM Thiazovivin.
iPSC Growth Medium: mTeSR™ Plus supplemented with 1% Penicillin/Streptomycin and 200 µg/ml Geneticin (G418).
iPSC Passage Medium: mTeSR™ Plus supplemented with 1% Penicillin/Streptomycin and 200 µg/ml Geneticin (G418) and 1 µM Thiazovivin.
2X Freezing Medium: 80% mTeSR™ Plus supplemented with 1% Penicillin/Streptomycin, 1 µM Thiazovivin and 20% DMSO (vol/vol).
Induction Media
iPSC Thaw Medium: mTeSR™ Plus supplemented with 1% Penicillin/Streptomycin (does NOT contain Thiazovivin) and doxycycline at various concentrations (0.5-1.5 µg/mL).
Applications
Study molecular pathways using temporally controlled induction of gene editing.
Generation of sgRNA-edited iPS cell pools.
Screening of pooled sgRNAs libraries to identify pluripotency, differentiation, and drug toxicity genetic modulators.
Shipping Temperature
-80°C
Notes
License Disclosure The iPSC technology is protected by several patents, including US patent Nos. 8048999, 8058065, 8129187, 8278104, 8530238, 8900871, 9404124, 9499797, 10519425, and patent pending, for which iPS Academia Japan, Inc. has been granted license rights with a sub-licensable right. The purchase of this cell line grants you a 10-year license to use this cell line in your immediate laboratory for internal research purposes. Commercial use of this cell line is not allowed. Commercial use requires the appropriate license from iPS Academia Japan, Inc. This cell line is for research use only, not for therapeutic or prophylactic use in humans or animals. Use in humans is strictly prohibited. This license does not permit you to share, distribute, sell, sublicense, or otherwise make the cell line available for use to other laboratories, departments, research institutions, hospitals, universities, or biotech companies. The license does not permit modification of the cell line in any way. Modification of this cell line or transfer to another facility of the cells requires a separate license or additional fees; contact [email protected] for details. Publications using this cell line should reference BPS Bioscience, Inc., San Diego. Inappropriate use or distribution of this cell line will result in revocation of the license and result in an immediate cease of sales and distribution of BPS products to your laboratory. BPS does not warrant the suitability of the cell line for any particular use and does not accept any liability in connection with the handling or use of the cell line.
Maintenance of the cells requires specific reagents such as specialty culture media, Matrigel™, Accutase™, RelesR™, and Thiazovivin that are not provided with the cells. Ensure that you have all reagents on hand prior to thawing the cells. Prepare media as indicated in section “Media Required for Cell Culture” below. Thiazovivin is a Rho Kinase inhibitor used to ensure that sensitive cell types such as iPS cells survive cell dissociation process and re-plate successfully. Thiazovivin is not stable in solution and should be added to the medium immediately before use.
Powered by Bioz
