کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1993410 1064665 2014 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
CRISPR/Cas9 mediated genome engineering in Drosophila
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
CRISPR/Cas9 mediated genome engineering in Drosophila
چکیده انگلیسی


• Targeted mutagenesis of Drosophila genes with CRISPR/Cas9 nucleases.
• Rapid production of novel synthetic guide RNAs by PCR and in vitro transcription.
• Simple and highly sensitive detection of mutations by high resolution melt analysis.
• Efficient generation of indels that can be transmitted through the germline.

Genome engineering has revolutionised genetic analysis in many organisms. Here we describe a simple and efficient technique to generate and detect novel mutations in desired target genes in Drosophila melanogaster. We target double strand breaks to specific sites within the genome by injecting mRNA encoding the Cas9 endonuclease and in vitro transcribed synthetic guide RNA into Drosophila embryos. The small insertion and deletion mutations that result from inefficient non-homologous end joining at this site are detected by high resolution melt analysis of whole flies and individual wings, allowing stable lines to be made within 1 month.

Figure optionsDownload as PowerPoint slide

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Methods - Volume 69, Issue 2, September 2014, Pages 128–136
نویسندگان
, ,