Efficient gene targeting in δCc.ku70 or δCc.lig4 mutants of the agaricomycete coprinopsis cinerea

Takehito Nakazawa, Yuki Ando, Kohei Kitaaki, Kiyoshi Nakahori, Takashi Kamada

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Coprinopsis cinerea is a model for studies of sexual development in agaricomycetes (homobasidiomycetes). Efficient gene targeting should facilitate such studies, especially because increasing genome and transcriptome information is now available in C. cinerea. To estimate the frequency of gene disruption by homologous integration in this fungus, we tried to disrupt Cc.wc-2, which encodes a WC-2 homolog, a partner of the fungal blue-light photoreceptor, WC-1. Disruption of Cc.wc-2 did not occur when recipients (protoplasts) of the disrupting construct were prepared from asexual spores, oidia, from the wild type, 326, while it occurred when protoplasts were prepared from mycelial cells from the same strain, albeit at a low frequency (3%). Double-stranded RNA-mediated silencing of a ku70 homolog, named Cc.ku70, or the lig4 homolog Cc.lig4 more or less increased the frequency of Cc.wc-2 targeting. On the basis of these results, we disrupted Cc.ku70 using a Cc.lig4-silenced strain. We then disrupted Cc.lig4 using the Cc.ku70 disruptant. We found that the disruption of Cc.ku70 or Cc.lig4 greatly enhanced gene targeting. In addition, this study demonstrates that Cc.wc-2 is involved in blue light perception in this fungus.

Original languageEnglish
Pages (from-to)939-946
Number of pages8
JournalFungal Genetics and Biology
Volume48
Issue number10
DOIs
Publication statusPublished - Oct 2011

Fingerprint

Gene Targeting
Protoplasts
Fungi
Light
Sexual Development
Double-Stranded RNA
RNA Interference
Spores
Transcriptome
Gene Frequency
Genome

Keywords

  • Cc.wc-2
  • Coprinus cinereus
  • Gene targeting
  • Homologous integration

ASJC Scopus subject areas

  • Genetics
  • Microbiology

Cite this

Efficient gene targeting in δCc.ku70 or δCc.lig4 mutants of the agaricomycete coprinopsis cinerea. / Nakazawa, Takehito; Ando, Yuki; Kitaaki, Kohei; Nakahori, Kiyoshi; Kamada, Takashi.

In: Fungal Genetics and Biology, Vol. 48, No. 10, 10.2011, p. 939-946.

Research output: Contribution to journalArticle

Nakazawa, Takehito ; Ando, Yuki ; Kitaaki, Kohei ; Nakahori, Kiyoshi ; Kamada, Takashi. / Efficient gene targeting in δCc.ku70 or δCc.lig4 mutants of the agaricomycete coprinopsis cinerea. In: Fungal Genetics and Biology. 2011 ; Vol. 48, No. 10. pp. 939-946.
@article{69afc1696dbd46429c058fdd112554dd,
title = "Efficient gene targeting in δCc.ku70 or δCc.lig4 mutants of the agaricomycete coprinopsis cinerea",
abstract = "Coprinopsis cinerea is a model for studies of sexual development in agaricomycetes (homobasidiomycetes). Efficient gene targeting should facilitate such studies, especially because increasing genome and transcriptome information is now available in C. cinerea. To estimate the frequency of gene disruption by homologous integration in this fungus, we tried to disrupt Cc.wc-2, which encodes a WC-2 homolog, a partner of the fungal blue-light photoreceptor, WC-1. Disruption of Cc.wc-2 did not occur when recipients (protoplasts) of the disrupting construct were prepared from asexual spores, oidia, from the wild type, 326, while it occurred when protoplasts were prepared from mycelial cells from the same strain, albeit at a low frequency (3{\%}). Double-stranded RNA-mediated silencing of a ku70 homolog, named Cc.ku70, or the lig4 homolog Cc.lig4 more or less increased the frequency of Cc.wc-2 targeting. On the basis of these results, we disrupted Cc.ku70 using a Cc.lig4-silenced strain. We then disrupted Cc.lig4 using the Cc.ku70 disruptant. We found that the disruption of Cc.ku70 or Cc.lig4 greatly enhanced gene targeting. In addition, this study demonstrates that Cc.wc-2 is involved in blue light perception in this fungus.",
keywords = "Cc.wc-2, Coprinus cinereus, Gene targeting, Homologous integration",
author = "Takehito Nakazawa and Yuki Ando and Kohei Kitaaki and Kiyoshi Nakahori and Takashi Kamada",
year = "2011",
month = "10",
doi = "10.1016/j.fgb.2011.06.003",
language = "English",
volume = "48",
pages = "939--946",
journal = "Fungal Genetics and Biology",
issn = "1087-1845",
publisher = "Academic Press Inc.",
number = "10",

}

TY - JOUR

T1 - Efficient gene targeting in δCc.ku70 or δCc.lig4 mutants of the agaricomycete coprinopsis cinerea

AU - Nakazawa, Takehito

AU - Ando, Yuki

AU - Kitaaki, Kohei

AU - Nakahori, Kiyoshi

AU - Kamada, Takashi

PY - 2011/10

Y1 - 2011/10

N2 - Coprinopsis cinerea is a model for studies of sexual development in agaricomycetes (homobasidiomycetes). Efficient gene targeting should facilitate such studies, especially because increasing genome and transcriptome information is now available in C. cinerea. To estimate the frequency of gene disruption by homologous integration in this fungus, we tried to disrupt Cc.wc-2, which encodes a WC-2 homolog, a partner of the fungal blue-light photoreceptor, WC-1. Disruption of Cc.wc-2 did not occur when recipients (protoplasts) of the disrupting construct were prepared from asexual spores, oidia, from the wild type, 326, while it occurred when protoplasts were prepared from mycelial cells from the same strain, albeit at a low frequency (3%). Double-stranded RNA-mediated silencing of a ku70 homolog, named Cc.ku70, or the lig4 homolog Cc.lig4 more or less increased the frequency of Cc.wc-2 targeting. On the basis of these results, we disrupted Cc.ku70 using a Cc.lig4-silenced strain. We then disrupted Cc.lig4 using the Cc.ku70 disruptant. We found that the disruption of Cc.ku70 or Cc.lig4 greatly enhanced gene targeting. In addition, this study demonstrates that Cc.wc-2 is involved in blue light perception in this fungus.

AB - Coprinopsis cinerea is a model for studies of sexual development in agaricomycetes (homobasidiomycetes). Efficient gene targeting should facilitate such studies, especially because increasing genome and transcriptome information is now available in C. cinerea. To estimate the frequency of gene disruption by homologous integration in this fungus, we tried to disrupt Cc.wc-2, which encodes a WC-2 homolog, a partner of the fungal blue-light photoreceptor, WC-1. Disruption of Cc.wc-2 did not occur when recipients (protoplasts) of the disrupting construct were prepared from asexual spores, oidia, from the wild type, 326, while it occurred when protoplasts were prepared from mycelial cells from the same strain, albeit at a low frequency (3%). Double-stranded RNA-mediated silencing of a ku70 homolog, named Cc.ku70, or the lig4 homolog Cc.lig4 more or less increased the frequency of Cc.wc-2 targeting. On the basis of these results, we disrupted Cc.ku70 using a Cc.lig4-silenced strain. We then disrupted Cc.lig4 using the Cc.ku70 disruptant. We found that the disruption of Cc.ku70 or Cc.lig4 greatly enhanced gene targeting. In addition, this study demonstrates that Cc.wc-2 is involved in blue light perception in this fungus.

KW - Cc.wc-2

KW - Coprinus cinereus

KW - Gene targeting

KW - Homologous integration

UR - http://www.scopus.com/inward/record.url?scp=80052261097&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80052261097&partnerID=8YFLogxK

U2 - 10.1016/j.fgb.2011.06.003

DO - 10.1016/j.fgb.2011.06.003

M3 - Article

VL - 48

SP - 939

EP - 946

JO - Fungal Genetics and Biology

JF - Fungal Genetics and Biology

SN - 1087-1845

IS - 10

ER -