Understanding the diversity of genetic outcomes from CRISPR-Cas generated homology-directed repair

Journal or Publishing Institution: Communications Biology

Date of Publication: 12/06/2019

Study: https://www.nature.com/articles/s42003-019-0705-y

Author(s): Sansbury, B.M., Hewes, A.M. and Kmiec, E.B.

Article Type: Peer Reviewed Study

Abstract:

As CRISPR-Cas systems advance toward clinical application, it is essential to identify all the outcomes of gene-editing activity in human cells. Reports highlighting the remarkable success of homology-directed repair (HDR) in the treatment of inherited diseases may inadvertently underreport the collateral activity of this remarkable technology. We are utilizing an in vitro gene-editing system in which a CRISPR-Cas complex provides the double-stranded cleavage and a mammalian cell-free extract provides the enzymatic activity to promote non-homologous end joining, micro-homology mediated end joining, and homology-directed repair. Here, we detail the broad spectrum of gene-editing reaction outcomes utilizing Cas9 and Cas12a in combination with single-stranded donor templates of the sense and nonsense polarity. This system offers the opportunity to see the range of outcomes of gene-editing reactions in an unbiased fashion, detailing the distribution of DNA repair outcomes as a function of a set of genetic tools.

Keywords: CRISPR-Cas, homology-directed repair (HDR), gene editing, plant breeding, genomic damage

Citation:

Sansbury, B.M., Hewes, A.M. and Kmiec, E.B., 2019. Understanding the diversity of genetic outcomes from CRISPR-Cas generated homology-directed repair. Communications Biology, 2(1), pp.1-10.

Category:

• Health effects

Record ID: 2522