PMID- 38528259
OWN - NLM
STAT- MEDLINE
DCOM- 20240327
LR  - 20240328
IS  - 1573-6822 (Electronic)
IS  - 0742-2091 (Print)
IS  - 0742-2091 (Linking)
VI  - 40
IP  - 1
DP  - 2024 Mar 26
TI  - Establishment and characterization of cytochrome P450 1A1 CRISPR/Cas9 Knockout 
      Bovine Foetal Hepatocyte Cell Line (BFH12).
PG  - 18
LID - 10.1007/s10565-024-09856-7 [doi]
LID - 18
AB  - The cytochrome P450 1A (CYP1A) subfamily of xenobiotic metabolizing enzymes 
      (XMEs) consists of two different isoforms, namely CYP1A1 and CYP1A2, which are 
      highly conserved among species. These two isoenzymes are involved in the 
      biotransformation of many endogenous compounds as well as in the bioactivation of 
      several xenobiotics into carcinogenic derivatives, thereby increasing the risk of 
      tumour development. Cattle (Bos taurus) are one of the most important 
      food-producing animal species, being a significant source of nutrition worldwide. 
      Despite daily exposure to xenobiotics, data on the contribution of CYP1A to 
      bovine hepatic metabolism are still scarce. The CRISPR/Cas9-mediated knockout 
      (KO) is a useful method for generating in vivo and in vitro models for studying 
      xenobiotic biotransformations. In this study, we applied the ribonucleoprotein 
      (RNP)-complex approach to successfully obtain the KO of CYP1A1 in a bovine foetal 
      hepatocyte cell line (BFH12). After clonal expansion and selection, CYP1A1 
      excision was confirmed at the DNA, mRNA and protein level. Therefore, RNA-seq 
      analysis revealed significant transcriptomic changes associated with cell cycle 
      regulation, proliferation, and detoxification processes as well as on iron, lipid 
      and mitochondrial homeostasis. Altogether, this study successfully generates a 
      new bovine CYP1A1 KO in vitro model, representing a valuable resource for 
      xenobiotic metabolism studies in this important farm animal species.
CI  - (c) 2024. The Author(s).
FAU - Iori, Silvia
AU  - Iori S
AD  - Department of Comparative Biomedicine and Food Science, University of Padua, 
      Viale Dell'Universita 16, Legnaro, 35020, Padua, Italy.
FAU - D'Onofrio, Caterina
AU  - D'Onofrio C
AD  - Department of Comparative Biomedicine and Food Science, University of Padua, 
      Viale Dell'Universita 16, Legnaro, 35020, Padua, Italy.
FAU - Laham-Karam, Nihay
AU  - Laham-Karam N
AD  - University of Eastern Finland, A.I. Virtanen Institute for Molecular Sciences, 
      Neulaniementie 2, 70211, Kuopio, Finland.
FAU - Mushimiyimana, Isidore
AU  - Mushimiyimana I
AD  - University of Eastern Finland, A.I. Virtanen Institute for Molecular Sciences, 
      Neulaniementie 2, 70211, Kuopio, Finland.
FAU - Lucatello, Lorena
AU  - Lucatello L
AD  - Department of Comparative Biomedicine and Food Science, University of Padua, 
      Viale Dell'Universita 16, Legnaro, 35020, Padua, Italy.
FAU - Lopparelli, Rosa Maria
AU  - Lopparelli RM
AD  - Department of Comparative Biomedicine and Food Science, University of Padua, 
      Viale Dell'Universita 16, Legnaro, 35020, Padua, Italy.
FAU - Gelain, Maria Elena
AU  - Gelain ME
AD  - Department of Comparative Biomedicine and Food Science, University of Padua, 
      Viale Dell'Universita 16, Legnaro, 35020, Padua, Italy.
FAU - Capolongo, Francesca
AU  - Capolongo F
AD  - Department of Comparative Biomedicine and Food Science, University of Padua, 
      Viale Dell'Universita 16, Legnaro, 35020, Padua, Italy.
FAU - Pauletto, Marianna
AU  - Pauletto M
AD  - Department of Comparative Biomedicine and Food Science, University of Padua, 
      Viale Dell'Universita 16, Legnaro, 35020, Padua, Italy.
FAU - Dacasto, Mauro
AU  - Dacasto M
AD  - Department of Comparative Biomedicine and Food Science, University of Padua, 
      Viale Dell'Universita 16, Legnaro, 35020, Padua, Italy.
FAU - Giantin, Mery
AU  - Giantin M
AD  - Department of Comparative Biomedicine and Food Science, University of Padua, 
      Viale Dell'Universita 16, Legnaro, 35020, Padua, Italy. mery.giantin@unipd.it.
LA  - eng
GR  - PhD fellowship/Universita degli Studi di Padova/
GR  - SID 2020-Prot. BIRD 207109/Universita degli Studi di Padova/
PT  - Journal Article
DEP - 20240326
PL  - Switzerland
TA  - Cell Biol Toxicol
JT  - Cell biology and toxicology
JID - 8506639
RN  - EC 1.14.14.1 (Cytochrome P-450 CYP1A1)
RN  - 0 (Xenobiotics)
RN  - 9035-51-2 (Cytochrome P-450 Enzyme System)
SB  - IM
MH  - Cattle
MH  - Animals
MH  - *Cytochrome P-450 CYP1A1/genetics/metabolism
MH  - *Xenobiotics
MH  - CRISPR-Cas Systems/genetics
MH  - Cytochrome P-450 Enzyme System/genetics/metabolism
MH  - Hepatocytes/metabolism
MH  - Cell Line
PMC - PMC10963470
OTO - NOTNLM
OT  - Bovine
OT  - CRISPR/Cas9
OT  - CYP1A1
OT  - Knockout
OT  - Liver cells
OT  - Transcriptome
COIS- The authors declare no competing interests.
EDAT- 2024/03/26 06:45
MHDA- 2024/03/27 06:43
PMCR- 2024/03/26
CRDT- 2024/03/26 00:28
PHST- 2023/12/23 00:00 [received]
PHST- 2024/03/21 00:00 [accepted]
PHST- 2024/03/27 06:43 [medline]
PHST- 2024/03/26 06:45 [pubmed]
PHST- 2024/03/26 00:28 [entrez]
PHST- 2024/03/26 00:00 [pmc-release]
AID - 10.1007/s10565-024-09856-7 [pii]
AID - 9856 [pii]
AID - 10.1007/s10565-024-09856-7 [doi]
PST - epublish
SO  - Cell Biol Toxicol. 2024 Mar 26;40(1):18. doi: 10.1007/s10565-024-09856-7.