Genome Editing - 4D-Nucleofector Visual

Genome editing enables the targeted modification of a cell’s DNA, making it a powerful tool for fundamental biological research, early stage drug discovery, and for the development of novel cellular therapeutics, e.g. immunotherapy.


The non-viral Nucleofector™ Technology from Lonza is a highly effective solution for ZFN, TALEN or CRISPR delivery across a wide range of cell types.


Technical Resources

Webinar: CRISPR - Tips and Tricks for Efficient Delivery

Webinar: CRISPR – Tips and Tricks for Efficient Delivery into Primary Cells and Cell Lines


Learn about the basic principle of CRISPR-based genome editing, and discuss best practices to achieve optimal delivery results for hard-to-transfect cell lines, primary cells or stem cells.


Register for the webinar >


Webinar: Validating and Scaling CRISPR Applications

Webinar: Validating and Scaling CRISPR Applications
Presented by GEN, sponsored by Lonza


Learn more about appropriate target validation techniques and scalable methodologies in the CRISPR genome-editing system.


Register to view recording >


Collection of white papers thumbnail 


White Papers and Technical Guides


Register and download these White Papers and Technical Guides on genome editing:


  • CRISPR/Cas9 White Paper: Comparing delivery of ribonucleoprotein (RNP) for targeted gene editing using Nucleofection
  • Genome Editing using Nucleofector™ Technology - a Technical Reference Guide: Containing a brief introduction to genome editing tools and technical tips for ZFN, TALEN or CRISPR delivery using the Nucleofector™ Technology


Register to download PDFs >


CRISPR/Cas 9 Cell Therapeutics Webinar presented by GEN



Webinar: CRISPR/Cas9 Cell Therapeutics –
The Next Generation of Cures
Presented by GEN, sponsored by Lonza

Topics include: opportunities and challenges for the use of CRISPR/Cas9 in gene/cell therapeutic applications and its impact on next-generation disease model systems and drug screening.


Register to view recording >

Introduction to Genome Editing 

Genome editing using engineered nucleases is a powerful tool to delete, insert or replace a gene at a targeted genomic location1. The nuclease is generating a double-strand break in the genomic DNA and thus inducing one of two possible cellular repair processes. Non-homologous end-joining (NHEJ) can lead to a deletion of the functional gene, since it is often accompanied with mutations when closing the break. If a partially homologous donor sequence is available, an insertion or replacement of a gene can take place via homology-dependent repair (HDR). To perform such a modification at a specific target sequence, the nuclease is fused to or interacts with sequence-specific DNA-binding component directing the nuclease to the target sequence.

Genome Editing Tools - Brief Overview

The typically used tools for site-specific genome editing tools are:

  • ZFN (Zinc Finger Nucleases)
  • TALEN (Transcriptional Activator-Like Effector Nucleases)
  • CRISPR/Cas9 (Clustered Regularly Interspaced Palindromic Repeats and CRISPR-Associated nuclease 9).

Nuclease Fok1 Fok1 Cas9
DNA binding molecule ZF Protein TALE protein GuideRNA (gRNA)
Type Fusion protein
→ High effort to modify for new targeting site
Fusion protein
→ High effort to modify for new targeting site
Protein + RNA
→ Easy to modify
→ Multiple targeting possible
Binding sites 2 sites (15 or 18 bp each)
→ High specificity
→ Low risk for off-target effects
2 sites (≥ 13 bp each)
→ High specificity
→ Low risk for off-target effects
1 site (18-20 bp + 3bp PAM)
→ Lower specificity
→ Higher risk for off-target effects



Genome Editing – Successful Transfection of CRISPR & Co. using Nucleofector™ Technology

All genome editing tools require co-transfer of several substrates into the cell type of interest for successful modification of genomic DNA. Lonza’s non-viral Nucleofector™ Technology has been shown to work as a reliable and sufficient method for transferring the required DNA- or RNA-based components into various cell lines and primary or stem cells, e.g., primary T cells, human embryonic stem cells (hESC) or induced pluripotent stem cells (iPSCs). 

  • High transfection efficiencies for a broad range of cell types, including iPSCs
  • Efficient co-transfection of various substrates
  • Same conditions for transfecting plasmids, DNA, mRNA or PCR cassettes, ssODN
  • Proven for ZFN, TALEN and CRISPR by more than 30 publications, including high ranking journals 

Besides using the Nucleofector™ Technology for genome editing in iPSCs or ESCs, it also offers a reliable method for iPSC generation.


If you need any technical guidance on using the Nucleofector™ Technology for genome editing contact Lonza Scientific Support.



1) Gaj T et al (2013) ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering. Trends in Biotechnology 31(7):397-405 (review)