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Products >  Stem_Cell_Research >  Gene_Editing >  Human_iPS_Cell_Editing_Systems

Start-To-Finish hiPS Cell Gene Editing and Single-Cell-Cloning Systems

The unique combination of precise, footprint-free gene editing using CRISPR/Cas9 and human induced pluripotent stem (hiPS) cells allows for a new level of sophistication in generating disease models to promote rapid advancement in the development of new therapeutics.

Our CRISPR/Cas9 system is a simple, RNA-programmable method to mediate genome editing in mammalian cells; it can be used to generate gene knockouts (via insertion/deletion) or knockins (via HDR) in hiPS cells. Once the Cas9-sgRNA ribonucleoprotein (RNP) complexes have been delivered, single cells must be isolated and expanded into clonal cell lines to isolate and screen for the genotype of interest. Traditionally, establishing clonal populations from edited hiPS cells grown and passaged as colonies is inefficient and time consuming; often, it results in cell death or premature differentiation. To overcome this challenge, we’ve developed complete systems that combine footprint-free gene editing via efficient delivery of Cas9-sgRNA RNP complexes and single-cell cloning of hiPS cells. Successful single-cell cloning is achieved using our monolayer-based culture system, which bypasses the challenges of colony-based culture by allowing single-cell passaging, promoting survival of clones, and supporting further clonal expansion.

If you already have edited hiPS cells and need to establish clones, choose:

  • The Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit (Cat. # Y30021). This is a complete system for efficient expansion and scale-up manufacturing of hiPS cells in a feeder-free and defined environment. The kit contains all the necessary reagents to go from seeding single hiPS cells into 96-well plates to expansion of clones into 48-well plates. Following clonal expansion, hiPS cells can continue to be cultured using the Cellartis DEF-CS 500 Culture System (Cat. # Y30010).

To generate your own edited clonal hiPS cell lines, try one of our complete systems containing a Cas9-sgRNA RNP complex delivery system (choose electroporation-based or gesicle-based delivery) and the Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit:

  • The Cellartis iPSC rCas9 Electroporation and Single-Cell Cloning System (Cat. # 632643) is a complete system that allows efficient gene editing of hiPS cells via electroporation, followed by single-cell cloning and expansion into 48-well plates. This system provides recombinant Cas9, as well as all the necessary reagents to produce high yields of sgRNAs, for electroporation into your cells. It also includes components from the Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit (discussed above) for performing single-cell cloning post-editing.
  • The Cellartis iPSC CRISPR/Cas9 Gesicle and Single-Cell Cloning System (Cat. # 632642) is a complete system that allows efficient gene editing of hiPS cells via gesicle delivery of Cas9-sgRNA RNP complexes, followed by single-cell cloning and expansion into 48-well plates. Gesicles are a nontoxic and efficient alternative to electroporation that does not rely on costly equipment or consumables. This system provides all the necessary reagents to produce these cell-derived nanovesicles, which deliver Cas9 and sgRNA to hiPS cells. It also includes components from the Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit (discussed above) for performing single-cell cloning post-editing.

  At-A-Glance   Documents   Images & Data   Resources

Features

  • Highly efficient gene editing in hiPS cellsuse either electroporation or gesicles to deliver Cas9 protein and sgRNA with no genomic integration and reduced off-target effects
  • Superior single-cell survivaledited hiPS cells exhibit high (typically ~50%) survival when seeded as single cells in a 96-well plate
  • Maintenance of pluripotency after editing and during single-cell cloninghiPS cells maintain high levels (>90%) of pluripotency markers Oct-4, TRA-1-60, and SSEA-4
  • Stable karyotype from start to finish—maintain a normal and stable karyotype throughout editing, single-cell cloning, and expansion
  • Flexibility to perform gene editing experiments your way—choose from complete kits that provide editing and single-cell cloning solutions using either electroporation-based (Cat. # 632643) or gesicle-based (Cat. # 632642) CRISPR/Cas9 techniques, or utilize the culture media kit (Cat. # Y30021) to perform efficient single-cell cloning following your own editing experiments
  • Continuous use of DEF-CS technology throughout gene editing/single-cell cloning experiments—use the Cellartis DEF-CS 500 Culture System (Cat. # Y30010) before and during gene editing experiments, then during scale-up after single-cell cloning experiments

Applications

  • CRISPR/Cas9-mediated gene editing of hiPS cells
  • Single-cell cloning of hiPS cells
  • Disease modeling

Additional Product Information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.


 
 
Products
Cat. # Product Contents Size Price License Units Select
Y30021 Cellartis® iPSC Single-Cell Cloning DEF-CS™ Culture Media Kit 1 Kit $555.00 License Statements
632642 Cellartis® iPSC CRISPR/Cas9 Gesicle and Single-Cell Cloning System 1 System $1,748.00 License Statements
632643 Cellartis® iPSC rCas9 Electroporation and Single-Cell Cloning System 1 System $1,689.00 License Statements
 

 

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