Knock-out applications with LentiFlash®

The CRISPR technology is revolutionizing the world of pharmaceutical research in many applications such as hereditary diseases, oncology or infectious diseases. This very promising tool, derived from a bacteria repair system, has a great potential for a large number of therapeutic applications.

However, for optimal use, this technology must be carried using an efficient delivery system.

An efficient delivery system implies:

  • A high penetration rate into target cells
  • A high level of editing of the targeted gene
  • The preservation of the viability and the original cell phenotype
  • An all-in-one delivery system (gRNA guide(s) + Cas9)
  • A transient Cas9 expression by mRNA delivery, since it has been demonstrated that compared to a long-term viral Cas9 expression, a transient Cas9 expression can substantially reduce off-target genome editing in cells(1,2).

With these needs in mind, Vectalys has created a new RNA delivery system named LentiFlash®. Through a fast and transitory RNA expression, this non-integrative particle is particularly suitable to carry CRISPR-Cas9 into any cell type while preserving their original cell phenotype and viability.

The data below, gathered by Vectalys and shown during the Fichier pdf AACR and Fichier pdf ASGCT annual meetings, demonstrate that the combination of LentiFlash® with the CRISPR-Cas9 technology can be applied to several therapeutic fields.

LentiFlash® carrying CRISPR-Cas9 for cancer immunotherapy:

T cell response inhibited by tumor cells
Interaction of T cells with Tumor cells after disruption of PD1 by LentiFlash™
Disruption of PD1 in primary T cells

Thanks to the knock-out of PD1, T cells will not be able to interact with PD-L1 and will be effective against the tumor. Therefore, this strategy is a promising approach for future immunotherapeutic programs.

LentiFlash® carrying CRISPR-Cas9 for antiviral strategies:

Disruption of CXCR4 in primary T cells

A safe cell engineering:

Safe cell engineering

Material & methods

Material and methods gene editing with LentiFlash

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  1. Hao Yin, Chun-Qing Song, Joseph R Dorkin, Lihua J Zhu, Yingxiang Li, Qiongqiong Wu, Angela Park, Junghoon Yang, Sneha Suresh, Aizhan Bizhanova, Ankit Gupta, Mehmet F Bolukbasi, Stephen Walsh, Roman L Bogorad, Guangping Gao, Zhiping Weng, Yizhou Dong, Victor Koteliansky, Scot A Wolfe, Robert Langer, Wen Xue, Daniel G Anderson. Therapeutic genome editing by combined viral and non-viral delivery of CRISPR system components in vivo. Nat Biotechnol. Author manuscript; available in PMC 2017 May 9. Published in final edited form as: Nat Biotechnol. 2016 Mar; 34(3): 328–333. Published online 2016 Feb 1. doi: 10.1038/nbt.3471
  2. Yin H, Kauffman KJ, Anderson DG. Delivery technologies for genome editing. Nat Rev Drug Discov. 2017 Jun;16(6):387-399. Epub 2017 Mar 24. Review. doi: 10.1038/nrd.2016.280.
  3. Laurence Zitvogel, Guido Kroemer. Targeting PD-1/PD-L1 interactions for cancer immunotherapy. Oncoimmunology. 2012 Nov 1; 1(8): 1223–1225. doi: 10.4161/onci.21335.