Genome Editing of Silkworms. 2017

Takuya Tsubota, and Hideki Sezutsu
Transgenic Silkworm Research Unit, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, 1-2 Owashi, Tsukuba, Ibaraki, 305-8634, Japan.

Silkworm is a lepidopteran insect that has been used as a model for a wide variety of biological studies. The microinjection technique is available and it is possible to cause transgenesis as well as target gene disruption via the genome editing technique. TALEN-mediated knock-out is especially effective in this species. We also succeeded in the precise and efficient integration of a donor vector using the Precise Integration into Target Chromosome (PITCh) method. Here, we describe protocols for ZFN, TALEN, and CRISPR/Cas9-mediated genome editing as well as the PITCh technique in the silkworm. We consider that all of these techniques can contribute to the further promotion of various biological studies in the silkworm and other insect species.

UI MeSH Term Description Entries
D008845 Microinjections The injection of very small amounts of fluid, often with the aid of a microscope and microsyringes. Microinjection
D000069896 Transcription Activator-Like Effector Nucleases Artificial nucleases that cleave DNA at a defined distance from specific DNA sequences recognized by TRANSCRIPTION ACTIVATOR-LIKE EFFECTORS. They are composed of an endodeoxyribonuclease fused to DNA-binding domains of the transcription activator-like effectors. TALEN,Nucleases, Transcription Activator-Like Effector,TALENs,Nucleases, Transcription Activator Like Effector,Transcription Activator Like Effector Nucleases
D000072669 Gene Editing Genetic engineering or molecular biology techniques that involve DNA REPAIR mechanisms for incorporating site-specific modifications into a cell's genome. Base Editing,Genome Editing,Editing, Base,Editing, Gene,Editing, Genome
D000075804 Zinc Finger Nucleases Genetically engineered nucleases that cleave DNA at a defined distance from specific DNA sequences recognized by ZINC FINGER DNA-BINDING DOMAINS. They are composed of a DNA cleaving domain adapted from DNA endonucleases fused to a zinc finger DNA-binding domain. Zinc Finger Nuclease,Zinc-Finger Nuclease,Zinc-Finger Nucleases,Finger Nuclease, Zinc,Finger Nucleases, Zinc,Nuclease, Zinc Finger,Nuclease, Zinc-Finger,Nucleases, Zinc Finger,Nucleases, Zinc-Finger
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012831 Bombyx A genus of silkworm MOTHS in the family Bombycidae of the order LEPIDOPTERA. The family contains a single species, Bombyx mori from the Greek for silkworm + mulberry tree (on which it feeds). A native of Asia, it is sometimes reared in this country. It has long been raised for its SILK and after centuries of domestication it probably does not exist in nature. It is used extensively in experimental GENETICS. (From Borror et al., An Introduction to the Study of Insects, 4th ed, p519) Bombyx mori,Silkmoths,Silkworms,Silkmoth,Silkworm,Bombyx morus,Bombyxs,mori, Bombyx
D049750 Genome, Insect The genetic complement of an insect (INSECTS) as represented in its DNA. Insect Genome,Genomes, Insect,Insect Genomes
D055879 Gene Knock-In Techniques Techniques used to add in exogenous gene sequence such as mutated genes; REPORTER GENES, to study mechanisms of gene expression; or regulatory control sequences, to study effects of temporal changes to GENE EXPRESSION. Gene Knock In,Gene Knock In Techniques,Gene Knock-In,Gene Knock Ins,Gene Knock-In Technique,Gene Knock-Ins,In, Gene Knock,Ins, Gene Knock,Knock In, Gene,Knock Ins, Gene,Knock-In Technique, Gene,Knock-In Techniques, Gene,Knock-In, Gene,Knock-Ins, Gene,Technique, Gene Knock-In,Techniques, Gene Knock-In
D064113 CRISPR-Cas Systems Adaptive antiviral defense mechanisms, in archaea and bacteria, based on DNA repeat arrays called CLUSTERED REGULARLY INTERSPACED SHORT PALINDROMIC REPEATS (CRISPR elements) that function in conjunction with CRISPR-ASSOCIATED PROTEINS (Cas proteins). Several types have been distinguished, including Type I, Type II, and Type III, based on signature motifs of CRISPR-ASSOCIATED PROTEINS. CRISPR Cas Systems,CRISPR-Cas System,System, CRISPR-Cas,Systems, CRISPR-Cas

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