Fast and sensitive detection of indels induced by precise gene targeting
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Fast and sensitive detection of indels induced by precise gene targeting. / Yang, Zhang; Steentoft, Catharina; Hauge, Camilla; Hansen, Lars; Thomsen, Allan Lind; Niola, Francesco; Vester-Christensen, Malene B; Frödin, Morten; Clausen, Henrik; Wandall, Hans H; Bennett, Eric P.
In: Nucleic Acids Research, Vol. 43, No. 9, e59, 2015, p. 1-8.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Fast and sensitive detection of indels induced by precise gene targeting
AU - Yang, Zhang
AU - Steentoft, Catharina
AU - Hauge, Camilla
AU - Hansen, Lars
AU - Thomsen, Allan Lind
AU - Niola, Francesco
AU - Vester-Christensen, Malene B
AU - Frödin, Morten
AU - Clausen, Henrik
AU - Wandall, Hans H
AU - Bennett, Eric P
N1 - © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.
PY - 2015
Y1 - 2015
N2 - 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.
AB - 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.
U2 - 10.1093/nar/gkv126
DO - 10.1093/nar/gkv126
M3 - Journal article
C2 - 25753669
VL - 43
SP - 1
EP - 8
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 9
M1 - e59
ER -
ID: 137197898