PMID- 30610625
OWN - NLM
STAT- MEDLINE
DCOM- 20190617
LR  - 20230311
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 1917
DP  - 2019
TI  - Rapid Screening of CRISPR/Cas9-Induced Mutants Using the ACT-PCR Method.
PG  - 27-32
LID - 10.1007/978-1-4939-8991-1_2 [doi]
AB  - The clustered regularly interspaced short palindromic repeats 
      (CRISPR)/CRISPR-associated protein 9 (Cas9) system provides a technological 
      breakthrough in targeted mutagenesis. However, a significant amount of time and 
      cost is required to screen for the CRISPR/Cas9-induced mutants from a typically 
      large number of initial samples. Here, we describe a cost-effective and sensitive 
      screening technique based on conventional polymerase chain reaction (PCR), termed 
      "annealing at critical temperature PCR" (ACT-PCR), for identifying mutants. 
      ACT-PCR requires only a single PCR step followed by agarose gel electrophoresis. 
      The simplicity of ACT-PCR makes it particularly suitable for rapid, large-scale 
      screening of CRISPR/Cas9-induced mutants.
FAU - Wang, Chun
AU  - Wang C
AD  - State Key Laboratory of Rice Biology, China National Rice Research Institute, 
      Chinese Academy of Agricultural Sciences, Hangzhou, China.
FAU - Wang, Kejian
AU  - Wang K
AD  - State Key Laboratory of Rice Biology, China National Rice Research Institute, 
      Chinese Academy of Agricultural Sciences, Hangzhou, China. wangkejian@caas.cn.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
SB  - IM
MH  - CRISPR-Cas Systems/genetics/physiology
MH  - Clustered Regularly Interspaced Short Palindromic Repeats/*genetics
MH  - Gene Editing/methods
MH  - Polymerase Chain Reaction/*methods
OTO - NOTNLM
OT  - ACT-PCR
OT  - CRISPR/Cas9
OT  - Cost-effective
OT  - Genome editing
OT  - Large-scale
OT  - Mutant screening
OT  - Rapid
EDAT- 2019/01/06 06:00
MHDA- 2019/06/18 06:00
CRDT- 2019/01/06 06:00
PHST- 2019/01/06 06:00 [entrez]
PHST- 2019/01/06 06:00 [pubmed]
PHST- 2019/06/18 06:00 [medline]
AID - 10.1007/978-1-4939-8991-1_2 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2019;1917:27-32. doi: 10.1007/978-1-4939-8991-1_2.