CC-BY-NC-ND 4.0 International license peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was not. ABSTRACT Recombineering, the use of endogenous homologous recombination systems to recombine DNA in vivo, is a commonly used technique for genome editing in microbes. Recombineering has not yet been developed for animals, where non-homolo...

Recombineering, the use of endogenous homologous recombination systems to recombine DNA in vivo, is a commonly used technique for genome editing in microbes. Recombineering has not yet been developed for animals, where non-homology-based mechanisms have been thought to dominate DNA repair. Here, we demonstrate, using Caenorhabditis elegans, that linear DNAs with short homologies (∼35 bases) eng...

Clustered regularly interspaced palindromic repeats (CRISPRs) and the CRISPR-associated (Cas) nuclease protect bacteria and archeae from foreign DNA by site-specific cleavage of incoming DNA. Type-II CRISPR-Cas systems, such as the Streptococcus pyogenes CRISPR-Cas9 system, can be adapted such that Cas9 can be guided to a user-defined site in the chromosome to introduce double-stranded breaks. ...

Shigella species (spp.) are gram-negative bacteria that are responsible for shigellosis. Although it can be controlled via antibiotics, increasing number of antibiotic resistant isolates of Shigella have been reported. Therefore, other strategies such as production of specific vaccine against this organism could be a suitable therapeutic approach. Attenuated live vaccines produced by gene delet...

Genome mosaicism in temperate bacterial viruses (bacteriophages) is so great that it obscures their phylogeny at the genome level. However, the precise molecular processes underlying this mosaicism are unknown. Illegitimate recombination has been proposed, but homeologous recombination could also be at play. To test this, we have measured the efficiency of homeologous recombination between dive...

A simple and efficient strategy for Bacterial Artificial Chromosome (BAC) recombineering based on co-selection is described. We show that it is possible to efficiently modify two positions of a BAC simultaneously by co-transformation of a single-stranded DNA oligo and a double-stranded selection cassette. The use of co-selection BAC recombineering reduces the DNA manipulation needed to make a c...

The retinal photoreceptors of a New World migratory bird, the bobolink (Dolichonyx oryzivorus) were examined using microspectrophotometry. Based on the absorbance spectra of their visual pigments and oil droplets, retinal photoreceptors include five classes of single cones, one double cone, and one rod. The single cones contain a long-wavelength pigment (lambda(max)=564nm) paired with a red or ...

shigella species (spp.) are gram-negative bacteria that are responsible for shigellosis. although it can be controlled via antibiotics, increasing number of antibiotic resistant isolates of shigella have been reported. therefore, other strategies such as production of specific vaccine against this organism could be a suitable therapeutic approach. attenuated live vaccines produced by gene delet...

Advances in DNA sequencing technology have facilitated the determination of hundreds of complete genome sequences both for bacteria and their bacteriophages. Some of these bacteria have well-developed and facile genetic systems for constructing mutants to determine gene function, and recombineering is a particularly effective tool. However, generally applicable methods for constructing defined ...