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March eNews

Research Update

Shorter Guide Strands Increase CRISPR/Cas Nuclease Specificity

Clustered, regularly interspaced, short palindromic repeat (CRISPR) RNA-guided nucleases have recently gained a lot of attention for their ability to manipulate the genome. These nucleases are particularly useful because they can target a specific genomic location through the use of guide RNA (gRNA). In nature, gRNA is approximately 100 nucleotides long. The specificity is determined by a 20 bp sequence ending with an NGG trinucleotide (5′-X(20)NGG-3′) that binds to the target RNA to be edited. gRNA tolerates up to 5 mismatches, particularly at the 5′ region. This can introduce unwanted insertion or deletion (indel) mutations, limiting the use of the CRISPR/Cas9 system. To combat this problem, Fu et al. attempt to improve the specificity through manipulation of the guide sequences in their manuscript entitled, Improving CRISPR-Cas nuclease specificity using truncated guide RNAs.

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Artificially Expanding DNA’s Genetic Code in Designer Microbes

Seemingly everyone these days knows about DNA and how seemingly simple pairings of A-T and G-C encode all life forms. Now, a team led by Floyd Romesberg, a biological chemist at the Scripps Research Institute in San Diego, California, has created a synthetic base pair to produce artificial DNA that replicates with six bases!

  • DNA isn’t What it Used To Be!
  • Scripps Team Demonstrates Six-Base DNA Replication in Living Bacteria
  • Are We On Our Way to Semi-synthetic Life-forms? With Vast Potential—But at What Risks?

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DNA-Day-OfferProf. Floyd E. Romesberg, Department of Chemistry, The Scripps Research Institute, La Jolla, California

RNA Guides for CRISPR Mediated Genome Editing

In the CRISPR system an RNA guide sequence, rather than a protein, directs targeting of the site of interest. The Cas9 protein is employed each time to perform the DNA cleavage.

Chemical synthesis of the RNA guide strand offers the opportunity to include chemical modifications that could influence the stability and specificity of the guide strand. In contrast to plasmid based CRISPR approaches, when a chemically synthesized guide strand is used in conjunction with Cas9 mRNA, there is no risk of DNA insertion. In addition to Cas9 mRNA, TriLink also offers guide strand synthesis. Go to OligoBuilder® to price and order your RNA guide strand today.

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A New Approach to Early Phase Therapeutic GMP Manufacturing

TriLink has been a key supplier of high quality research grade and diagnostic grade GMP material since 1996, and will now offers therapeutic GMP production. Our state-of-the-art 2,000 sq ft GMP manufacturing facility will be fully operational July 2014.

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Previous Blog Post

Modified mRNA Mania
  • Biosynthetic modified mRNA for gene-based therapy without the gene!
  • AstraZeneca bets up to $420M on Moderna’s “messenger RNA therapeutics”
  • “Me-too” Pharma frenzy to follow?

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