Long-read sequencing for fast and robust identification of correct genome-edited alleles: PCR-based and Cas9 capture methods.

ABSTRACT:

Background

Recent developments in CRISPR/Cas9 genome-editing tools have facilitated the introduction of precise alleles, including genetic intervals spanning several kilobases, directly into the embryo. However, the introduction of donor templates, via homology directed repair, can be erroneous or incomplete and these techniques often produce mosaic founder animals. Thus, newly generated alleles must be verified at the sequence level across the targeted locus. Screening for the presence of the desired mutant allele using traditional sequencing methods can be challenging due to the size of the interval to be sequenced, together with the mosaic nature of founders.

Methodology/principal findings

In order to help disentangle the genetic complexity of these animals, we tested the

SUBMITTER: McCabe CV 

PROVIDER: S-EPMC10954187 | biostudies-literature | 2024 Mar

REPOSITORIES: biostudies-literature