iPSC Derivation and Gene Editing Facility
Our first class facility provides stem cell derivation and gene-editing services to researchers worldwide.
About us
This facility brings together a team of highly experienced stem cell and molecular biologists with over 40 years of combined experience to generate standard and gene-edited iPSC lines in feeder-free and chemically defined medium in a manner that is both rapid and cost-effective.
A unique feature of our core is the implementation of a streamlined one-step gene-editing/reprogramming protocol that utilises episomal reprogramming vectors and the Cas9/CRISPR gene-editing system to facilitate the rapid and efficient generation of clonally derived gene-edited iPSC lines from primary fibroblasts or blood.
We also have the capacity to perform gene-editing in already existing human pluripotent stem cell lines as well as the generation of lineage-specific reporter lines.
Upon initial contact, we liaise with customers to devise the ideal strategy for achieving a successful gene-editing outcome based upon the ever-expanding number of gene-editing tools available including:
- SpCas9
- Cas12
- ABE and CBE Base Editing
- Prime Editing
The iPSC Derivation & Gene Editing Core also offers other key services such as teratoma assays, MEF production, and customised training.
Order form
iPSC Derivation & Gene Editing Core Facility Order Form314.06 KB
Contact us
For more information, please contact us:
Email:
show email address
Phone: +61 3 9936 6456 or +61 3 9936 6164
Meet the team
Jia Yi Kuah
Our services
Generation of patient iPS cell lines
Service includes:
- Episomal or Sendai delivery of reprograming factors
- From fibroblast, peripheral/cord blood cells and PBMCs
- iPSCs generated can be cultured to the requirements of the end user (on MEFs or MEF-free)
- All supplied iPSCs (up to three clones) will be checked by molecular karyotype and expression of pluripotency markers and confirmed mycoplasma negative
- Provision of a standard report
CRISPR/Cas9-mediated gene knock-out
Service includes:
- Consultation and generation/validation of sgRNA plasmid(s)
- Electroporation of sgRNA and Cas9 mRNA into nominated fibroblast or hPSC line
- PCR screening to identify successfully modified clones
- DNA Sanger sequencing to verify mutant clones
- Expansion of up to 3 mutant clones
- Freezing (4 vials per line)
- SNP array to assess genomic integrity of at least 2 lines
- *Upon request, extra lines can be assessed by SNP array (@ $335 per line + 10% GST for external clients)
At completion, the user will be provided with sgRNA plasmid maps, reports with PCR analysis and Sanger sequencing files confirming successfully modified lines, SNP analyses, and mycoplasma-free certification.
CRISPR/Cas9-mediated gene knock-in or gene repair
Service includes:
- Consultation and generation/validation of sgRNA plasmid(s)
- Mycoplasma testing (if cells are provided)
- Electroporation of sgRNA, Cas9 mRNA and homologous template into nominated fibroblast or hPSC line
- PCR screening to identify successfully modified clones
- DNA Sanger sequencing to verify mutant clones
- Expansion of up to 3 successfully modified clones. Additional matched isogenic controls will also be provided in the case of gene repair experiments.
- Freezing (4 vials per line)
- SNP array to assess genomic integrity of at least 2 lines
*Upon request, extra lines can be assessed by SNP array (@ $335 per line + 10% GST for external clients)
At completion, the user will be provided with: sgRNA plasmid maps, reports with PCR analysis, Sanger sequencing files confirming successfully modified lines and SNP analyses, and mycoplasma-free certification.
Blood processing and cryopreservation
- Isolation of peripheral blood mononuclear cells (PBMCs) from whole blood in EDTA/Heparin/Citrate tubes stored at RT
- Cryopreservation
- Short-term storage in LN2 when requesting generation of iPSCs
Culture reagents for the maintenance of iPSC lines
Preparation and provision of basic medias and mouse embryonic feeder cells (MEFs)
Teratomas
- Supply of mice and preparation of cell lines to be injected
- Subcutaneous injection of undifferentiated cells
- Subcutaneous injection/implant of differentiated cells is also available
- Monitoring and teratoma detection
- Teratoma harvest and provision to researcher
hES/iPS cell culture service
Services include but are not limited to:
- Clone derivation
- Cell maintenance
- Cell expansion for cryopreservation
MEF and MEF-free options are available
Cell lines must test negative for mycoplasma (documentation required) prior to entering the facility.
Training & support
We offer:
- Customised one-on-one training which is complimentary for researchers that use our iPSC derivation and/or gene-editing services.
- Specific training around iPSC maintenance and passaging using various formats (feeder and feeder-free methods).
- Cryopreservation and revival of iPSCs. Ongoing advisory and technical assistance covering all aspects of culture.
Disclaimers
If the investigator supplies primary fibroblasts, they can be provided as frozen vials or as live cultures. Cells must test negative for mycoplasma (documentation required) prior to the commencement of reprogramming/gene-editing experiments. Primary fibroblasts should be at a low passage (ideally <10) and should be actively proliferating. Senescent cultures are not amenable to reprogramming. In some situations, we may not obtain successfully modified lines from the first round. In this scenario, a second round will be attempted using a modified strategy (e.g., second sgRNA and/or different starting cell line). The investigator will be charged 30% for two failed attempts.
For primary fibroblast lines that reprogram poorly using the simultaneous reprogramming/gene editing approach, iPSC lines will be generated first and then subsequently gene-edited to obtain enough clones to screen. In these instances, we will charge the equivalent of gene editing an already established iPSC line, i.e., $14,000 ($15,400 including GST).
iPSC Derivation & Gene Editing Proces diagram
Downloading the MCRI iPSC Derivation & Gene Editing Proces diagram (png, 621.24 KB)
Pricing
Price does not include shipment of cell lines on dry ice which can be organised with your choice or courier.
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Service/Product
|
Internal Clients
|
External Clients (inclusive of 10% GST)
|
|---|---|---|
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iPSC derivation from skin or blood (Episomal or Sendai available)
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Starting at $3000
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Starting at $3,300
|
|
Gene knock-out
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$6000
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$6600
|
|
Point mutation repair/knock-in
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$12,000-$14,000
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$13,200-$15,400
|
|
Reporter gene knock-in
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$14,000
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$15,400
|
|
Bulk cell expansion and cryopreservation
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$200 for 4 vials
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$220 for 4 vials
|
|
Blood processing (PBMC isolation and cryopreservation)
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$200 per sample
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$220 per sample
|
|
Teratoma assay
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$550 per mouse
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$605 per mouse
|
|
MEFs and MEF-culture specific medias for the maintenance of iPSCs
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Upon request
|
Upon request
|
|
Training and support
|
Please enquire
|
Please Enquire
|
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