Fast and sensitive detection of indels induced by precise gene targeting

Zhang Yang, Catharina Steentoft, Camilla Hauge, Lars Hansen, Allan Lind Thomsen, Francesco Niola, Malene Bech Vester-Christensen, Morten Frödin, Henrik Clausen, Hans H. Wandall, Eric P. Bennett

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Abstract

The nuclease-based gene editing tools are rapidly transforming capabilities for altering the genome of cells and organisms with great precision and in high throughput studies. A major limitation in application of precise gene editing lies in lack of sensitive and fast methods to detect and characterize the induced DNA changes. Precise gene editing induces double-stranded DNA breaks that are repaired by error-prone non-homologous end joining leading to introduction of insertions and deletions (indels) at the target site. These indels are often small and difficult and laborious to detect by traditional methods. Here we present a method for fast, sensitive and simple indel detection that accurately defines indel sizes down to ±1 bp. The method coined IDAA for Indel Detection by Amplicon Analysis is based on tri-primer amplicon labelling and DNA capillary electrophoresis detection, and IDAA is amenable for high throughput analysis.
Original languageEnglish
Article numbere59
JournalNucleic Acids Research
Volume43
Issue number9
Number of pages8
ISSN0305-1048
DOIs
Publication statusPublished - 2015

Bibliographical note

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which
permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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