RRC ID 47714
Author Yoshida K, Hozumi A, Treen N, Sakuma T, Yamamoto T, Shirae-Kurabayashi M, Sasakura Y.
Title Germ cell regeneration-mediated, enhanced mutagenesis in the ascidian Ciona intestinalis reveals flexible germ cell formation from different somatic cells.
Journal Dev Biol
Abstract The ascidian Ciona intestinalis has a high regeneration capacity that enables the regeneration of artificially removed primordial germ cells (PGCs) from somatic cells. We utilized PGC regeneration to establish efficient methods of germ line mutagenesis with transcription activator-like effector nucleases (TALENs). When PGCs were artificially removed from animals in which a TALEN pair was expressed, somatic cells harboring mutations in the target gene were converted into germ cells, this germ cell population exhibited higher mutation rates than animals not subjected to PGC removal. PGC regeneration enables us to use TALEN expression vectors of specific somatic tissues for germ cell mutagenesis. Unexpectedly, cis elements for epidermis, neural tissue and muscle could be used for germ cell mutagenesis, indicating there are multiple sources of regenerated PGCs, suggesting a flexibility of differentiated Ciona somatic cells to regain totipotency. Sperm and eggs of a single hermaphroditic, PGC regenerated animal typically have different mutations, suggesting they arise from different cells. PGCs can be generated from somatic cells even though the maternal PGCs are not removed, suggesting that the PGC regeneration is not solely an artificial event but could have an endogenous function in Ciona. This study provides a technical innovation in the genome-editing methods, including easy establishment of mutant lines. Moreover, this study suggests cellular mechanisms and the potential evolutionary significance of PGC regeneration in Ciona.
Volume 423(2)
Pages 111-125
Published 2017-3-15
DOI 10.1016/j.ydbio.2017.01.022
PII S0012-1606(16)30698-4
PMID 28161521
MeSH Animals Animals, Genetically Modified Base Sequence Ciona intestinalis / cytology* Ciona intestinalis / genetics* Electroporation Gene Knockout Techniques Genome Germ Cells / cytology* Germ Cells / metabolism Homeodomain Proteins / genetics Homeodomain Proteins / metabolism Intestinal Mucosa / metabolism Intestines / embryology Male Metamorphosis, Biological Mutagenesis / genetics* Mutation / genetics Mutation Rate Organ Specificity Ovum / cytology Regeneration* Spermatozoa / cytology Tail Transcription Activator-Like Effector Nucleases / metabolism
IF 2.896
Times Cited 10
C.intestinalis / (O.japonicus) Tg[MiCiTnIG]2 Tg[MiCiTnIG]3 Tg[MiCiNutG]3 Tg[MiCiNutK]4 Tg[MiCiNutK]8 EJ[MiTSAdTPOG]75 EJ[MiTSAdTPOG]87 EJ[MiTSAdTPOG]101 Wild C. int