PMID- 36370349
OWN - NLM
STAT- MEDLINE
DCOM- 20221115
LR  - 20221115
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2582
DP  - 2023
TI  - Protocol for CRISPR/Cas Genome Editing for Investigating Cell Communication 
      Network.
PG  - 157-167
LID - 10.1007/978-1-0716-2744-0_11 [doi]
AB  - The Cellular Communication Network Factor (CCN) family is composed of six 
      members: CCN1/CYR61, CCN2/CTGF, CCN3/NOV, CCN4/WISP1, CCN5/WISP2, and CCN6/WISP3. 
      The second member, CCN2/CTGF is a matricellular protein that promotes 
      extracellular matrix (ECM) synthesis and controls angiogenesis. On the other 
      hand, moonlighting/matrix metalloproteinase 3 (MMP3) is an ECM-degrading enzyme 
      that also functions as an intracellular transcription factor. Importantly, 
      extracellular MMP3 is uptaken into cells, translocating into nuclei, and 
      transcriptionally activating CCN2/CTGF gene in cancer and chondrocytes. Thus, the 
      MMP3-CTGF axis balances the matrix metabolism and turnover in the tissue and 
      tumor microenvironments. We established an MMP3 knockout cell line using the 
      CRISPR/Cas9 system, demonstrating the sequential regulatory events of the 
      MMP3-CCN2 axis in the microenvironment. Notably, our protocol is useful for 
      generation of CCN knockout cells as well. Here we serve a protocol of the 
      CRISPR/Cas9-based gene targeting in cultured cells for investigating cellular 
      communication network.
CI  - (c) 2023. The Author(s), under exclusive license to Springer Science+Business 
      Media, LLC, part of Springer Nature.
FAU - Okusha, Yuka
AU  - Okusha Y
AD  - Division of Molecular and Cellular Biology, Department of Radiation Oncology, 
      Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.
FAU - Eguchi, Takanori
AU  - Eguchi T
AD  - Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and 
      Pharmaceutical Sciences, Okayama University, Okayama, Japan. 
      eguchi@okayama-u.ac.jp.
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
RN  - EC 3.4.24.17 (Matrix Metalloproteinase 3)
RN  - 139568-91-5 (Connective Tissue Growth Factor)
SB  - IM
MH  - *Matrix Metalloproteinase 3/genetics/metabolism
MH  - *Gene Editing
MH  - CRISPR-Cas Systems/genetics
MH  - Gene Expression Regulation
MH  - Cell Communication/genetics
MH  - Connective Tissue Growth Factor/metabolism
OTO - NOTNLM
OT  - CCN2/CTGF
OT  - CRISPR/Cas9
OT  - cellular communication network
OT  - exosomes
OT  - genetic knockout
OT  - genome editing
OT  - moonlighting/matrix metalloproteinase
OT  - tissue microenvironment
EDAT- 2022/11/13 06:00
MHDA- 2022/11/16 06:00
CRDT- 2022/11/12 11:21
PHST- 2022/11/12 11:21 [entrez]
PHST- 2022/11/13 06:00 [pubmed]
PHST- 2022/11/16 06:00 [medline]
AID - 10.1007/978-1-0716-2744-0_11 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2023;2582:157-167. doi: 10.1007/978-1-0716-2744-0_11.