Home
Resources
Publications
Pharmacological Inhibition of DNA-PK Stimulates Cas9-mediated Genome Editing

Pharmacological Inhibition of DNA-PK Stimulates Cas9-mediated Genome Editing

Francis Robert, Mathilde Barbeau, Sylvain Éthier, Josée Dostie, Jerry Pelletier

Genome Med. 2015 Aug 27;7(1):93.

PMID: 26307031

Abstract:

Background:
The ability to modify the genome of any cell at a precise location has drastically improved with the recent discovery and implementation of CRISPR/Cas9 editing technology. However, the capacity to introduce specific directed changes at given loci is hampered by the fact that the major cellular repair pathway that occurs following Cas9-mediated DNA cleavage is the erroneous non-homologous end joining (NHEJ) pathway. Homology-directed recombination (HDR) is far less efficient than NHEJ and makes screening of clones containing directed changes time-consuming and labor-intensive.
Methods:
We investigated the possibility of pharmacologically inhibiting DNA-PKcs, a key player in NHEJ, using small molecule inhibitors (NU7441 and KU-0060648), to ameliorate the rates of HDR repair events. These compounds were tested in a sensitive reporter assay capable of simultaneously informing on NHEJ and HDR, as well as on an endogenous gene targeted by Cas9.
Results:
We find that NU7441 and KU-0060648 reduce the frequency of NHEJ while increasing the rate of HDR following Cas9-mediated DNA cleavage.
Conclusions:
Our results identify two small molecules compatible for use with Cas9-editing technology to improve the frequency of HDR.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP881375004	KU-0060648		881375-00-4	Price

As a specialist in analytical chemical Products, Alfa Chemistry offers consumers a wealth of raw materials and a full range of technologies and services.

QUICK LINKS

HEADQUARTERS

Tel:

Fax:

Email: