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Biology & Genetics · 2012

A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity

Martin Jinek, Krzysztof Chylinski, Ines Fonfara, Michael Hauer, Jennifer A. Doudna, Emmanuelle Charpentier

University of California, Berkeley · Umeå University

Science, vol. 337, pp. 816–821 ·

Cited by 15,000+Open access
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This paper turned a bacterial immune system into a programmable gene-editing tool. Doudna, Charpentier and colleagues showed that the Cas9 enzyme, guided by a single engineered RNA, can be directed to cut any chosen DNA sequence.

Founded CRISPR-Cas9 genome editing; won the 2020 Nobel Prize in Chemistry.

  • Cas9 uses two RNAs (crRNA and tracrRNA) to recognise and cut target DNA at a specified site.
  • The two RNAs can be fused into a single 'guide RNA' that fully programs Cas9.
  • Changing the guide sequence retargets the cut — a simple, universal editing system.

Biochemical reconstitution: purified Cas9 protein and synthetic RNAs were tested in vitro for site-specific cleavage of DNA, with mutational analysis of the enzyme's two nuclease domains and engineering of a chimeric single-guide RNA.

Keywords

Biology & Genetics

Initial Sequencing and Analysis of the Human Genome

Lander et al. · 2001 · Nature

The public Human Genome Project's first analysis of the human genome sequence. It presented a draft covering most of the genome and drew early conclusions about the surprisingly small number of human genes.

Cited by 24,000+Open access

Biology & Genetics

Viable Offspring Derived from Fetal and Adult Mammalian Cells

Wilmut et al. · 1997 · Nature

The paper announcing Dolly the sheep, the first mammal cloned from an adult cell. It proved that the nucleus of a fully differentiated cell can be reset to support the development of a whole new animal.

Cited by 6,000+

Étude Science indexes and summarises this work; it is not the publisher. The summary above is written by Étude. For the definitive text, figures, and data, please consult the original publication via the link above. Jinek et al. (2012) hold the rights to the original work.