High-Efficiency Gene Editing

High-Efficiency Gene Editing Service in Donor-Derived CD34+ HSPCs

Expanding on the Cooperative Center of Excellence in Hematology (CCEH) at Fred Hutchinson Cancer Center’s expertise in providing Human CD34+ HSPCs, we have established a high-efficiency and high-viability protocol for gene editing in CD34+ cells. We are offering a Gene Editing Service (Basic) in conjunction with provision of donor-derived CD34+ HSPCs. Using non-viral delivery of Cas9:sgRNA ribonucleoprotein complexes, we generate high efficiency biallelic single, double and triple gene Knock Outs (KO) in human CD34+ cells. On a case-by-case basis we are also providing custom editing services not limited to but covering the use of alternative Cas systems, targeted insertion and library screens.
 

Validation of Gene Knock Out in CD34+ Cells 

indel formation chart

Indel formation

Consistent indel formation and KO efficiency is detected across donors.

Protein Expression chart

Protein Expression

Robust loss of protein expression is detected following gene editing.

Erythroid Differentiation graph

Erythroid Differentiation

Penetrant phenotypes following KO are observed during in vitro erythroid differentiation.

Data from Kuppers et al. 2023

Overview of Base Gene Editing Service

Overview of base gene editing service

We've optimized this technique to minimize time in culture, and, as a result, cells maintain multi-lineage potential in vitro and in vivo.

1. Consultation
2. Design of sgRNAs
3. Selection of appropriate controls
4. KO procedure
5. Sequencing-based validation of targeting event
6. Provision of viably frozen KO CD34+ cells along with donor-matched controls cells

Please send a service request through the contact link below and we will get back to you about scheduling a consultation.
 

In addition to the services covered in the basic service, custom editing services will provide closer collaboration to tailor editing to your experimental needs. What is covered will vary on a case-by-case basis but may include things such as alternative Cas systems, targeted insertion, library screens or additional protocol development and optimization. Please send us a description of your project through the link below and we will get back to you about our ability to support your needs and to schedule a consultation.

Base Gene Editing Service

  • Initial new gene KO or new control setup/validation fee: $700
  • Additional new genes at the same time: $400
  • 1st Donor — 1 million KO CD34+ cells and 1 million Control CD34+ cells  $2,100
  • Additional Donors Edited at the Same Time (up to 2 additional) — 1 million KO CD34+ cells and 1 million Control CD34+ cells $850 per donor
  • Each additional 1 million cells (Either KO or Control) — up to 9 million additional cells of each $300

     
  • FedEx Next-Day on Dry Ice
  • Other shipping options can be accommodated for an additional expense.
     

Publications and Protocols

We are available to discuss our services or answer your questions.

More Information

Contact Us

Cell Manipulation Tools Core
Fred Hutchinson Cancer Center C3-175
P.O. Box 19024
Seattle, WA  98109-1024

Work Location: Hutchinson Building, C3-175
Daniel Kuppers, PhD

Daniel Kuppers, PhD

Co-Director, CCEH Lab
Patrick Paddison, PhD

Patrick Paddison, PhD

Director, CCEH Lab