What paper outlines the discovery of how the CRIPER DNA, CrRNA, and Cas9 interact?


2 Answers 2


The two "classic" papers cited in terms of Crispr/Cas9 discovery include the Doudna/Charpentier initial discovery, and the Feng Zhang application to gene editing. There is an ongoing, famous patent dispute as to whom really discovered the Crispr/Cas9 gene editing technology.

As a quick timeline that better answers your question, a good publication is Cell Paper and there is actually a great podcast, but I will summarize:

  1. Yoshizumi Ishino et al. in 1987 Discovers Crispr mechanism - the authors simply noticed the odd regular repeats of DNA in bacteria and noted: “no sequence homologous to these has been found elsewhere in procaryotes, and the biological significance of these sequences is not known.”

  2. Francisco Mojica in 1993-2005 - Furthers research into CRISPR, at this time the terms has been coined by Jansen, however it is Mojica who really notices the Crispr pattern in several bacterium and suggests that it is involved with an immune response.

  3. Alexander Bolotin in 2005 - Looking into the bacteria Streptococcus thermophilus, Bolotin discovers that the genetic material in between the spacers is viral DNA, providing definitive evidence of the Crispr hypothesis.

  4. Marakova in 2006 - First proposal that the protein Cas9 can be used in a bacterial adaptive immune response, although Bolotin is also credited around this time from noticing the use of Cas9.

  5. Koonin in 2006 - The first one to piece the Crispr storage of viral DNA and the Cas9 reports of an immune response together as a related system that might be able to use the Crispr DNA with Cas9 in an immune system.

  6. Marraffini in 2007 - Studying the Crispr-Cas9 system further, he provides the first definitive microbiological evidence that Cas9 is a programmable restriction enzyme, and that it targets DNA not RNA. At the same time Philippe Horvath, Danisco France also provided the best evidence yet of the adaptive immunity system.

  7. Doudna and Carpentier in 2010 - Through sequencing, definitively put the Crispr and Cas9 system together as complete unit that is able to cut DNA, they also described tracRNA as the RNA piece that join with Cas9 to program DNA cutting.

  8. Feng Zhang in 2013 - While Doudna previously suggested that the Cas9 system could be used to target mammalian DNA, Zhang was the first one to actually that it was possible and therefore open to the door as the Crispr/Casa9 system as a tool that easily edit genomes.

While I definitley missed several key steps, and did not fully explain the actual mechanism of Crispr/Cas9, I think this provides a decent timeline. For more information about the actual mechanism see: Harvard, Wired, Science News.


This paper will tell you about it.https://www.ncbi.nlm.nih.gov/pubmed/?term=A+Programable+Dual-RNA-Guided+Endonuclease+In+Adaptive+Bacterial+Immunity

I am not allowed to seee the full text,but there it is.

Here is a full text link of that paper http://science.sciencemag.org/content/337/6096/816.full


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