PMID- 31523770
OWN - NLM
STAT- MEDLINE
DCOM- 20201217
LR  - 20201217
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2069
DP  - 2020
TI  - Genetic Manipulation of MRSA Using CRISPR/Cas9 Technology.
PG  - 113-124
LID - 10.1007/978-1-4939-9849-4_9 [doi]
AB  - The clustered regularly interspersed short palindromic repeat (CRISPR)/Cas9 
      system has emerged as an efficient genome engineering method attributed to its 
      high efficiency and versatility. By generating a lethal double-strand DNA break 
      in the target genome, the CRISPR/Cas9 system is capable of selecting the 
      separated crossover events occurring in the traditional genetic manipulation 
      methods in one step, therefore enabling rapid and efficient genome editing in 
      Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA). By 
      engineering the fusion of a cytidine deaminase APOBEC1 and a Cas9 nickase, a base 
      editor was further developed as a highly efficient gene inactivation and point 
      mutation tool in S. aureus. Here we describe a detailed protocol for 
      CRISPR/Cas9-based genome editing in S. aureus, including genome modification and 
      base editing. This protocol outlines in detail the design of primers, the 
      construction and transformation of editing plasmids, as well as the verification 
      of sequence-specific CRISPR/Cas9-mediated mutagenesis in S. aureus.
FAU - Chen, Weizhong
AU  - Chen W
AD  - School of Physical Science and Technology, ShanghaiTech University, Shanghai, 
      China.
FAU - Ji, Quanjiang
AU  - Ji Q
AD  - School of Physical Science and Technology, ShanghaiTech University, Shanghai, 
      China. quanjiangji@shanghaitech.edu.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
MH  - *Gene Editing
MH  - Methicillin-Resistant Staphylococcus aureus/*genetics
MH  - *Mutagenesis
MH  - Plasmids/*genetics
MH  - *Point Mutation
OTO - NOTNLM
OT  - Base editing
OT  - CRISPR/Cas9
OT  - Genome editing
OT  - MRSA
OT  - Staphylococcus aureus
OT  - pCasSA
OT  - pnCasSA-BEC
EDAT- 2019/09/17 06:00
MHDA- 2020/12/18 06:00
CRDT- 2019/09/17 06:00
PHST- 2019/09/17 06:00 [entrez]
PHST- 2019/09/17 06:00 [pubmed]
PHST- 2020/12/18 06:00 [medline]
AID - 10.1007/978-1-4939-9849-4_9 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2020;2069:113-124. doi: 10.1007/978-1-4939-9849-4_9.