Fast and Sensitive Detection of Indels Induced by Precise Gene Targeting

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2015 Mar 11

PMID 25753669

Citations 96

Authors

Zhang Yang

Catharina Steentoft

Camilla Hauge

Lars Hansen

Allan Lind Thomsen

Francesco Niola

Malene B Vester-Christensen

Morten Frodin

Henrik Clausen

Hans H Wandall

Eric P Bennett

Affiliations

Soon will be listed here.

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.

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