Efficient CRISPR/Cas9 Plasmids for Rapid and Versatile Genome Editing in Drosophila | G3: Genes | Genomes | Genetics
The CRISPR-associated Cas9 endonuclease can recognize and cleave target DNA in combination with a single chimeric guide RNA ( ... The use of single-stranded oligodeoxynucleotides (ssODN) as HDR donors is a simple, yet powerful, approach to introduce precise ... 2012 Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria. Proc. Natl. Acad. ... 2012 A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337: 816-821. ...
A Dual sgRNA Approach for Functional Genomics in Arabidopsis thaliana | G3: Genes | Genomes | Genetics
2012 A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337: 816-821. doi:10.1126/science. ... that binds and protects single stranded DNA (Khodaverdian et al. 2017). Moreover, the TIDE method used in our study limits ... prepared genomic DNA and genotyped using Cas9 specific primers to identify null-segregants (Figure 3A). The genomic DNA of ... A generic nuclease, Cas9, is guided to a target DNA sequence (protospacer) by associating with an artificial single guide RNA ( ...
Integrated Analysis of miRNA and mRNA Expression Profiles Reveals Functional miRNA-Targets in Development Testes of Small Tail...
pmirGLO was digested by restriction endonuclease SacI and XhoI. pmirGLO and target gene fragments were ligated by T4 DNA ligase ... Total RNA was extracted from testis tissues using Trizol Reagent (Invitrogen, USA). RNA integrity number (RIN) and RNA quality ... Specific primers were designed and synthesized by Sangon Biotech (Shanghai, China); primer information is shown in Table 1. ... Then, the short mRNA fragments were reverse-transcribed and amplified to synthetize double-stranded cDNA. Emulsion PCR was ...
Assessment of a Split Homing Based Gene Drive for Efficient Knockout of Multiple Genes | G3: Genes | Genomes | Genetics
The GDe contains two guide RNAs (gRNAs) targeting the DNA cleavage at white and yellow loci, and an eye-specific marker (3xP3- ... 2012 A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science 337: 816-821. doi:10.1126/science. ... Single-fly genomic DNA preps were prepared by homogenizing a fly in 30µl of a freshly prepared squishing buffer (10mM Tris-Cl ... double stranded DNA breaks (DSBs) on the opposite chromosome can result in the drive allele being used as a template (i.e., ...
Iron Deprivation in Synechocystis: Inference of Pathways, Non-coding RNAs, and Regulatory Elements from Comprehensive...
Single-stranded RNA probes were generated by in vitro transcription from amplicons containing the T7 promoter as previously ... Activated Fur attaches to specific DNA sequences (termed Fur boxes) in the promoter regions of iron-acquisition genes and ... a homing endonuclease within the tRNAfMet group I intron (slr0915), and two proteins of unknown function (slr1634 and slr0376 ... Hybridization was carried out with 1.65 µg RNA per array, according to the Agilent instructions for 4 × 44 k single-color ...
Meeting Report: The Allied Genetics Conference 2016 | G3: Genes | Genomes | Genetics
... unwinding the double helix to allow the RNA to bind and form an RNA/DNA hybrid that displaces the other DNA strand. Finally, ... It could even be simplified; Doudnas RNA expertise enabled her to connect the two RNA molecules and create one single RNA ... 2012 A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337(6096): 816-821. ... These methods also introduce double-strand breaks at specific sequences, triggering DNA repair via either the nonhomologous end ...