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CRISPR-Cas genome editing technology

16/06/2021

Nominated by: Spanish Bioindustry Association – AseBio, Lithianian Biotechnology Association

 

People in nomination:

  1. Professor Francisco Martinez Mojica, Department of Physiology, Genetics and Microbiology, University of Alicante, Spain
  2. Dr. Virginijus Šikšnys, Life Sciences Center, Vilnius University, Lithuania

In a nutshell: CRISPR is a family of DNA sequences found within the genomes of microorganisms such as bacteria and archaea. These sequences are fragments of bacteriophage genomes that previously infected the cell. They protect the bacteria from future infections, essentially operating as an acquired immunity. CRISPR are found in approximately 50% of sequenced bacterial genomes.

Cas are enzymes that use the CRISPR sequences to find and cleave DNA associated with those sequences.

CRISPR is a guide and Cas is the editing tool, creating a complex through which DNA can be precisely edited.

The history of CRISPR-Cas stretches back to the 1980s, with scientific observations and advances from researchers around the world helping to build understanding and applications. This is the story of global learning that contributed to arguably one of the most significant new technologies of the 21st century so far.

Europe was a key player in the development of a genome editing technique that has scooped a Nobel Prize and is now fuelling product developments across health, food, feed and agriculture.

CRISPR first found a name in 2002, through the work of Professor Francisco Martinez Mojica, who was investigating an odd detail he had detected in an archaea, Haloferax mediterranei, on the Santa Pola salt flats. After years of considering a series of genetic sequences repeating at regular intervals in the genome of microorganisms, he gave these sequences the name of CRISPR, standing for “Clustered Regularly Interspaced Short Palindromic Repeats.”

Mojica discovered that between these repeat sequences lay fragments of viral DNA, a remnant of the bacteria’s past invaders. This lightbulb moment, showed the microbial CRISPR to be a vaccine against previously encountered aggressors, a defensive memory transmitted from generation to generation of microorganisms. He suggested that CRISPR systems in procaryotes, with their different repeats and interspace sequences, are part of a sophisticated defence system. His seminal results were published in 2005.

Work published in Utrecht, provided knowledge to add the Cas side of the story, that the repeat cluster was accompanied by a set of four genes that make up the CRISPR Associated Systems, or Cas, although the exact mode of action was not yet determined.

These scientific discoveries were critical to lay the ground on which modern genome editing techniques were developed.

In 2011, Professor Dr Virginijus Šikšnys added to the story, demonstrating that the CRISPR-Cas9 system could be transferred from one bacterium to another and the Cas9 protein could be used for programmed genome editing, contributing to the understanding of the structure and function of restriction enzymes. He was amongst the first scientists to demonstrate programmable DNA cleavage by the Cas9 protein.

In the same period, alongside work from the Šikšnys laboratory, understanding of how CRISPR-Cas works was also advanced through the work of eventual Nobel prize winners Jennifer Doudna and Emmanuelle Charpentier, who described in vitro the compounds and mode of action of the CRISPR-Cas9 system of the bacteria Streptococcus pyogenes that could find and cleave a DNA target.  The Nobel Prize for Chemistry was awarded in 2020 to both of them for discovering “a method of genome editing”, demonstrating again that breakthrough science is the work of decades and multiple scientists across the world.

In 2018, Šikšnys, together with Doudna and Charpentier were joint recipients of the Kavli Prize in Nanoscience “for the invention of CRISPR-Cas9, a precise nanotool for editing DNA, causing a revolution in biology, agriculture, and medicine”.

A tool such as CRISPR-Cas has opened the door to applications into gene editing across the spectrum.

Food production: As far back as 2005, applications of the as yet undetermined mechanism was discovered in research into yoghurt development.  It is now being applied into food and farming to develop pro-biotic cultures and immunised bacterial production systems against infection.

Crop production: Applications within agriculture are a generational advance on original genetic modification of plants, able to advance traits such as drought tolerance and nutritional profile in response to climate change and farming needs, in addition to improving biofuel production.

Healthcare: CRISPR has been applied into multiple disease areas as scientists target unmet clinical needs. Diseases such as HIV and sickle cell disease have been directly targeted, while mosquitos could be modified to prevent transmission of malaria.  CRISPR is a valuable diagnostic tool as well, enabling understanding of gene disease relationships, which would then allow development of treatments.

References

• Identification of genes that are associated with DNA repeats in prokaryotes
• Intervening Sequences of Regularly Spaced Prokaryotic Repeats Derive from Foreign Genetic Elements. 
• Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria. 
• 2018 Kavli Prize for NanoScience
• 2016 Warren Alpert Prize

 

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Alexandra Simionca
Alexandra Simionca
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