Screening Methods to Identify TALEN-Mediated Knockout Mice
スポンサーリンク
概要
- 論文の詳細を見る
Genome editing with site-specific nucleases, such as zinc-finger nucleases or transcription activator-like effector nucleases (TALENs), and RNA-guided nucleases, such as the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated) system, is becoming the new standard for targeted genome modification in various organisms. Application of these techniques to the manufacture of knockout mice would be greatly aided by simple and easy methods for genotyping of mutant and wild-type pups among litters. However, there are no detailed or comparative reports concerning the identification of mutant mice generated using genome editing technologies. Here, we genotyped TALEN-derived enhanced green fluorescent protein (eGFP) knockout mice using a combination of approaches, including fluorescence observation, heteroduplex mobility assay, restriction fragment length polymorphism analysis and DNA sequencing. The detection sensitivities for TALEN-induced mutations differed among these methods, and we therefore concluded that combinatorial testing is necessary for the screening and determination of mutant genotypes. Since the analytical methods tested can be carried out without specialized equipment, costly reagents and/or sophisticated protocols, our report should be of interest to a broad range of researchers who are considering the application of genome editing technologies in various organisms.
- Japanese Association for Laboratory Animal Scienceの論文
Japanese Association for Laboratory Animal Science | 論文
- F344の亜系統, F344/DuとF344/Nの下顎骨の形態計測学的比較
- 高血圧高脂血症自然発症ラットにおける高フィブリノーゲン血症と高血圧および高脂血症合併モデルとしての有用性
- 大腸菌および緑膿菌由来エンドトキシンが心血行動態に及ぼす影響
- Pax6遺伝子をヘミ欠失したマウスの消化器系腫瘍
- サイクロオキシゲナーゼ-2(COX-2)とトランスフォーミング成長因子-β1(TGF-β1)に対する免疫反応性を示すリチャードソンジリス (Spermophilus richardsonii) の頬腺由来自然発生性腺癌の一例