RRC ID 77173
Author Sasakura Y, Horie T.
Title Improved Genome Editing in the Ascidian Ciona with CRISPR/Cas9 and TALEN.
Journal Methods Mol Biol
Abstract The ascidian Ciona intestinalis type A (or Ciona robusta) is an important organism for elucidating the mechanisms that make the chordate body plan. CRISPR/Cas9 and TAL effector nuclease (TALEN) are widely used to quickly address genetic functions in Ciona. Our previously reported method of CRISPR/Cas9-mediated mutagenesis in this animal has inferior mutation rates compared to those of TALENs. We here describe an updated way to effectively mutate genes with CRISPR/Cas9 in Ciona. Although the construction of TALENs is much more laborious than that of CRISPR/Cas9, this technique is useful for tissue-specific knockouts that are not easy even by the optimized CRISPR/Cas9 method.
Volume 2637
Pages 375-388
Published 2023-1-1
DOI 10.1007/978-1-0716-3016-7_28
PMID 36773161
MeSH Animals CRISPR-Cas Systems / genetics Ciona* / metabolism Ciona intestinalis* / genetics Ciona intestinalis* / metabolism Gene Editing / methods Gene Knockout Techniques Transcription Activator-Like Effector Nucleases / genetics Transcription Activator-Like Effector Nucleases / metabolism Transcription Activator-Like Effectors / genetics
C.intestinalis / (O.japonicus) Wild C. int pHTB-TALEN-NG::2A::mCherry pHTB-TALEN-NI::2A::mCherry pHTB-TALEN-NN::2A::mCherry pHTB-TALEN-HD::2A::mCherry pEF1a>TALEN-NG::2A::mCherry pEF1a>TALEN-NI::2A::mCherry pEF1a>TALEN-NN::2A::mCherry pEF1a>TALEN-HD::2A::mCherry