Harnessing innovation to drive discoveries

Quick Order|Contact us| CUSTOMER LOGIN

888.266.5066 (Toll Free)|Fax: 650.968.2277

PrecisionX™ HR Targeting Vectors

Genome engineering with
homology-directed recombination vectors

HR Targeting Vectors

  • Dual Marker Options
  • Built-in Insulators
  • LoxP Sites for Removal
  • HR-arm Multiple Cloning Sites

PrecisionX™ HR Targeting Vectors

   

Recent advances in tools available for precise genome engineering of target cells have revolutionized the field of biology. Transcriptional activator- like effector nucleases (TALENs), first described in the plant pathogen Xanthomonas sp., have shown that researchers can efficiently target any genomic DNA sequence using a pair of custom TALEN proteins whose DNA-binding modules can recognize individual DNA nucleotides based on an elegant amino acid cipher (Boch et al. 2009). Based on the explosion of recent publications that have successfully demonstrated the utility of the TALEN platform in many eukaryotic model organisms ranging from zebrafish to humans, it has quickly established itself as the de facto platform for genome engineering.

The Cas9/CRISPR technology is based on a bacterial system for combating invading viral and nucleic acid sequences. Originally discovered in the pathogenic bacterium Streptococcus pyogenes, this system uses an unique series of short RNAs (collectively termed as "guide RNA") to specifically target a complementary DNA sequence, and upon binding, leads to recruitment of an endonuclease called Cas9 to specifically induce a double-stranded break (DSB) in the DNA sequence targeted by the guide RNA (Bhaya et al. 2011, Jinek et al. 2012). Similar to TALENs, the introduction of DSBs in the DNA leads to recruitment of the cellular repair machinery to drive non-homologous end-joining (NHEJ) processes in absence of homologous sequences, whose error-prone nature often leads to formation of indels in the targeted DNA sequences.

However, in order to fully leverage the powerful nature of these genome engineering platforms, there needs to be a set of ancillary tools that will allow detection of cells that have been modified by TALEN or Cas9 systems. The majority of genome editing events affect only a percentage of the total population of transfected cells, estimated at 1-80% for mono- or bi-allelic modifications depending on the platform, the cell type, and the DNA target of interest. Due to the wide variation in activity, performing downstream phenotypic assays in a background of wild-type cells is challenging, especially if the phenotype in question is subtle or difficult to discern. Therefore, tools in the form of donor or targeting vectors that contain 1) fluorescent or antibiotic selection markers and 2) a gene fragment of interest to knock-in, knock-out, or correct a wild-type sequence will be extremely useful for obtaining a homogeneous population of cells whose genomes have been successfully targeted. SBI has generated a suite of homologous recombination targeting vectors suitable for the purpose of engineering both protein coding and non-coding genes including microRNAs and LncRNAs genomic loci.