SEP 17, 2020 8:00 AM PDT

Scalable Manufacturing and Nanovesicle Delivery of CRISPR-Cas9 Ribonucleoproteins Using a cGMP- Compliant Cell Engineering Platform

Sponsored by: MaxCyte
Speaker

Abstract

DATE: September 17, 2020

TIME: 8:00am PT

 

CRISPR-Cas9 has tremendous potential as a therapeutic tool for treating human diseases. However, prolonged expression of the nuclease and gRNA from viral vectors in an in vivo context may cause unwanted off-target activity and immunogenicity. To overcome these safety issues, a system was recently developed for transient delivery of CRISPR-Cas9 ribonucleoprotein (RNP), recruiting Cas9 protein by chemically-induced dimerization and sgRNA via a viral RNA packaging signal into extracellular nanovesicles.  This system, termed NanoMEDIC (nanomembrane-derived extracellular vesicles for the delivery of macromolecular cargo), demonstrates efficient genome editing in various hard-to-transfect cell types, including human induced pluripotent stem (iPS) cells and myoblasts, and also in vivo in a luciferase reporter mouse model. To demonstrate the feasibility of industrial scale manufacturing, a xeno-free suspension culture system to produce NanoMEDIC by flow electroporation was developed using chemically defined media. We show that electroporation parameters using a cGMP-compliant cell engineering platform could be optimized to increase NanoMEDIC production in HEK293 cells, and the process is scalable for flow electroporation of over one billion cells. These findings indicate that NanoMEDIC manufacturing may be amenable for large scale production.

 

Learning objectives:

  • Identify considerations for switching from an adherent cell lipofection method to suspension cell electroporation 

  • Assess experimental parameters that can be optimized for efficient electroporation of HEK293 cells  

  • Demonstrate scalable extracellular nanoparticle production by electroporation 

 

Webinars will be available for unlimited on-demand viewing after live event

 

LabRoots is approved as a provider of continuing education programs in the clinical laboratory sciences by the ASCLS P.A.C.E. ® Program. By attending this webinar, you can earn 1 Continuing Education credit once you have viewed the webinar in its entirety.


Show Resources
You May Also Like
FEB 15, 2023 7:00 AM PST
FEB 15, 2023 7:00 AM PST
Date: February 15, 2023 Time: 7:00am (PST), 10:00pm (EST), 4:00pm (CET) While not all microscopy samples can fluoresce, all can scatter light, and this scattered light can be imaged. This ha...
OCT 13, 2022 9:00 AM +08
OCT 13, 2022 9:00 AM +08
First Broadcast: Date: October 12, 2022 Time: 8:00am PDT, 11:00am EDT Second Broadcast: Date: October 12, 2022 Time: 9:00am SGT The new Embedded CryoSPARC Live, now fully integrated with t...
DEC 07, 2022 9:00 AM PST
C.E. CREDITS
DEC 07, 2022 9:00 AM PST
Date: December 07, 2022 Time: 9:00am (PST), 12:00pm (EST), 6:00pm (CET) Join us for an exciting live panel webinar, as we celebrate the 60th anniversary of Gibco Cell Culture. The panelists...
AUG 10, 2022 10:00 AM PDT
AUG 10, 2022 10:00 AM PDT
Date: August 10, 2022 Time: 10:00am PDT, 1:00pm EDT The global pandemic has increased focus and scrutiny on molecular diagnostic assay development, resulting in a need for assays that provid...
AUG 24, 2022 7:00 AM PDT
AUG 24, 2022 7:00 AM PDT
Date: August 24, 2022 Time: 7:00am (PDT), 10:00pm (EDT), 4:00pm (CEST) Light field microscopy was first introduced in 2006, and allows users to capture the 4D light field within the microsco...
NOV 09, 2022 8:00 AM PST
C.E. CREDITS
NOV 09, 2022 8:00 AM PST
Date: November 09, 2022 Time: 8:00am (PST), 11:00am (EST), 5:00pm (CET) The field of cell and gene therapy is rapidly growing. In particular, the use of lentiviruses in CAR-T applications is...
Loading Comments...
Show Resources