Heads of Laboratories
Investigator, Howard Hughes Medical Institute
Laboratory of Bacteriology
CRISPR-Cas systems enable bacteria and other microbes to acquire immunity against viruses by capturing snippets of their DNA. Marraffini investigates the molecular mechanisms that make CRISPR immunity possible, as well as its evolutionary implications. His lab also explores genome editing and other potential applications for CRISPR-Cas systems.
Sequence-directed genetic interference pathways control gene expression and preserve genome integrity in all kingdoms of life. In many bacteria and most archaea, CRISPRs—clustered, regularly interspaced, short palindromic repeats—specify a recently discovered genetic interference pathway that protects cells from phages and conjugative plasmids. Within CRISPR sequences, the repeats are separated by short spacer sequences that match phage or plasmid genomes and specify the targets of interference.
Spacer sequences are transcribed into CRISPR RNAs (crRNAs)—small RNAs that, through base-pairing interactions with the target sequence, guide Cas nucleases to the invasive nucleic acid. Upon infection, CRISPR arrays can acquire new repeat-spacer units that match the infecting phage or plasmid. In this way, CRISPR-Cas systems provide adaptive and inheritable immunity to the bacterial cell. The spacer content of CRISPR arrays reflects the many different invaders encountered by the host and can be expanded rapidly in response to new ones. Accordingly, CRISPR loci constitute a form of genetic memory that ensures the rejection of new, returning, and ever-present invading DNA molecules.
Marraffini uses Staphylococcus epidermidis and Streptococcus pyogenes as model systems for studying CRISPR immunity. The clinical isolate S. epidermidis RP62a harbors a CRISPR spacer that matches the nickase gene (nes) that is present in nearly all staphylococcal conjugative plasmids and prevents their spread. Using this system, Marraffini revealed that the CRISPR-Cas machinery targets DNA, rather than RNA, directly. Work in the Marraffini lab also demonstrated that the S. pyogenes crRNA-guided Cas9 DNA nuclease constitute a formidable tool for genetic engineering.
Marraffini’s current research employs molecular genetic and biochemical approaches to analyze the genesis and function of CRISPR-Cas systems. He ultimately hopes to answer fundamental questions about how CRISPR-Cas systems destroy their targets, how the genetic memory is generated, and how CRISPR-Cas immunity affects the evolution of bacteria and archaea.
Marraffini is a faculty member in the David Rockefeller Graduate Program, the Tri-Institutional M.D.-Ph.D. Program, and the Tri-Institutional Ph.D. Program in Chemical Biology.
Lic. in biotechnology, 1998
University of Rosario
Ph.D. in microbiology, 2007
University of Chicago
Northwestern University, 2008–2010
Assistant Professor, 2010–2016
Associate Professor 2016–2018
The Rockefeller University
Howard Hughes Medical Institute
RNA Society Award, 2010
Searle Scholar, 2011
Rita Allen Foundation Scholar, 2012
NIH Director’s New Innovator Award, 2012
The Rockefeller University Distinguished Teaching Award, 2013
40 Under 40, Cell, 2014
Hans Sigrist Prize, 2015
Earl and Thressa Stadtman Scholar Award, 2016
Howard Hughes Medical Institute Faculty Scholar, 2016
Albany Medical Center Prize in Medicine and Biomedical Research, 2017
Gabrielle H. Reem and Herbert J. Kayden Early-Career Innovation Award, 2017
NIH Director’s Pioneer Award, 2017
Samai, P. et al. Co-transcriptional DNA and RNA cleavage during Type III CRISPR-Cas immunity. Cell 161, 1164–1174 (2015).
Heler, R. et al. Cas9 specifies functional viral targets during CRISPR-Cas adaptation. Nature 519, 199–202 (2015).
Goldberg, G.W. et al. Conditional tolerance of temperate phages via transcription-dependent CRISPR-Cas targeting. Nature 514, 633–637 (2014).
Cong, L. et al. Multiplex genome engineering using CRISPR/Cas systems. Science 339, 819–823 (2013).
Jiang, W. et al. RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nat. Biotechnol. 31, 233–239 (2013).
Dr. Marraffini is a faculty member in the David Rockefeller Graduate Program, the Tri-Institutional M.D.-Ph.D. Program, and the Tri-Institutional Ph.D. Program in Chemical Biology.