PMID- 37529831
OWN - NLM
STAT- MEDLINE
DCOM- 20230913
LR  - 20230927
IS  - 1940-3372 (Electronic)
IS  - 1940-3372 (Linking)
VI  - 16
IP  - 3
DP  - 2023 Sep
TI  - Temporally gene knockout using heat shock-inducible genome-editing system in 
      plants.
PG  - e20376
LID - 10.1002/tpg2.20376 [doi]
AB  - Clustered regularly interspaced short palindromic repeat 
      (CRISPR)/CRISPR-associated nuclease 9 (Cas9) has emerged as a powerful tool to 
      generate targeted loss-of-function mutations for functional genomic studies. As a 
      next step, tools to generate genome modifications in a spatially and temporally 
      precise manner will enable researchers to further dissect gene function. Here, we 
      present two heat shock-inducible genome-editing (IGE) systems that efficiently 
      edit target genes when the system is induced, thus allowing us to target specific 
      developmental stages. For this conditional editing system, we chose the natural 
      heat-inducible promoter from heat-shock protein 18.2 (HSP18.2) from Arabidopsis 
      thaliana and the synthetic heat-inducible promoter heat shock-response element 
      HSE-COR15A to drive the expression of Cas9. We tested these two IGE systems in 
      Arabidopsis using cyclic or continuous heat-shock treatments at the seedling and 
      bolting stages. A real-time quantitative polymerase chain reaction analysis 
      revealed that the HSP18.2 IGE system exhibited higher Cas9 expression levels than 
      the HSE-COR15A IGE system upon both cyclic and continuous treatments. By 
      targeting brassinosteroid-insensitive 1 (BRI1) and phytoene desaturase (PDS), we 
      demonstrate that both cyclic and continuous heat inductions successfully 
      activated the HSP18.2 IGE system at the two developmental stages, resulting in 
      highly efficient targeted mutagenesis and clear phenotypic outcomes. By contrast, 
      the HSE-COR15A IGE system was only induced at the seedling stage and was less 
      effective than the HSP18.2 IGE system in terms of mutagenesis frequencies. The 
      presented heat shock-IGE systems can be conditionally induced to efficiently 
      inactivate genes at any developmental stage and are uniquely suited for the 
      dissection and systematic characterization of essential genes.
CI  - (c) 2023 The Authors. The Plant Genome published by Wiley Periodicals LLC on behalf 
      of Crop Science Society of America.
FAU - Liang, Zhen
AU  - Liang Z
AUID- ORCID: 0000-0002-5242-4644
AD  - School of Life Science, Shanxi University, Taiyuan, Shanxi, China.
FAU - Wei, Sha
AU  - Wei S
AD  - School of Life Science, Shanxi University, Taiyuan, Shanxi, China.
FAU - Wu, Yuqing
AU  - Wu Y
AD  - School of Life Science, Shanxi University, Taiyuan, Shanxi, China.
FAU - Guo, Yingjie
AU  - Guo Y
AD  - Research Institute of Big Data Science and Industry, Shanxi University, Taiyuan, 
      Shanxi, China.
FAU - Zhang, Ben
AU  - Zhang B
AD  - School of Life Science, Shanxi University, Taiyuan, Shanxi, China.
FAU - Yang, Honghu
AU  - Yang H
AD  - School of Life Science, Shanxi University, Taiyuan, Shanxi, China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20230802
PL  - United States
TA  - Plant Genome
JT  - The plant genome
JID - 101273919
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Gene Knockout Techniques
MH  - *CRISPR-Cas Systems
MH  - *Genome, Plant
MH  - Heat-Shock Response
MH  - Immunoglobulin E/genetics
EDAT- 2023/08/02 06:42
MHDA- 2023/09/13 06:41
CRDT- 2023/08/02 04:03
PHST- 2023/06/22 00:00 [revised]
PHST- 2023/04/20 00:00 [received]
PHST- 2023/07/23 00:00 [accepted]
PHST- 2023/09/13 06:41 [medline]
PHST- 2023/08/02 06:42 [pubmed]
PHST- 2023/08/02 04:03 [entrez]
AID - 10.1002/tpg2.20376 [doi]
PST - ppublish
SO  - Plant Genome. 2023 Sep;16(3):e20376. doi: 10.1002/tpg2.20376. Epub 2023 Aug 2.