Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems

DiCarlo, James E.; Norville, Julie E.; Mali, Prashant; Rios, Xavier; Aach, John; Church, George M.

doi:10.1093/nar/gkt135

Title: Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems

Journal Article · Mon Mar 04 00:00:00 EST 2013 · Nucleic Acids Research

DOI:https://doi.org/10.1093/nar/gkt135· OSTI ID:1904692

DiCarlo, James E. ^[1]; Norville, Julie E. ^[2]; Mali, Prashant ^[2]; Rios, Xavier ^[2]; Aach, John ^[2]; Church, George M. ^[2]

Boston University, MA (United States); Harvard Medical School, Boston, MA (United States)
Harvard Medical School, Boston, MA (United States)

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated (Cas) systems in bacteria and archaea use RNA-guided nuclease activity to provide adaptive immunity against invading foreign nucleic acids. Here, we report the use of type II bacterial CRISPR-Cas system in Saccharomyces cerevisiae for genome engineering. The CRISPR-Cas components, Cas9 gene and a designer genome targeting CRISPR guide RNA (gRNA), show robust and specific RNA-guided endonuclease activity at targeted endogenous genomic loci in yeast. Using constitutive Cas9 expression and a transient gRNA cassette, we show that targeted double-strand breaks can increase homologous recombination rates of single- and double-stranded oligonucleotide donors by 5-fold and 130-fold, respectively. In addition, co-transformation of a gRNA plasmid and a donor DNA in cells constitutively expressing Cas9 resulted in near 100% donor DNA recombination frequency. Our approach provides foundations for a simple and powerful genome engineering tool for site-specific mutagenesis and allelic replacement in yeast.

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Research Organization:: Harvard Univ., Cambridge, MA (United States)

Sponsoring Organization:: USDOE Office of Science (SC); National Science Foundation (NSF); National Institutes of Health (NIH)

Grant/Contract Number:: FG02-02ER63445; SA5283-11210; P50 HG005550

OSTI ID:: 1904692

Journal Information:: Nucleic Acids Research, Vol. 41, Issue 7; ISSN 0305-1048

Publisher:: Oxford University PressCopyright Statement

Country of Publication:: United States

Language:: English

References (21)

New Methods Enabling Efficient Incorporation of Unnatural Amino Acids in Yeast Wang, Qian; Wang, Lei Journal of the American Chemical Society, Vol. 130, Issue 19, p. 6066-6067 https://doi.org/10.1021/ja800894n	journal	May 2008
Fidelity of Mitotic Double-Strand-Break Repair in Saccharomyces cerevisiae: A Role for SAE2 / COM1 Rattray, Alison J.; McGill, Carolyn B.; Shafer, Brenda K. Genetics, Vol. 158, Issue 1 https://doi.org/10.1093/genetics/158.1.109	journal	May 2001
Chromosomal site-specific double-strand breaks are efficiently targeted for repair by oligonucleotides in yeast Storici, F.; Durham, C. L.; Gordenin, D. A. Proceedings of the National Academy of Sciences, Vol. 100, Issue 25 https://doi.org/10.1073/pnas.2036296100	journal	November 2003
A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity Jinek, M.; Chylinski, K.; Fonfara, I. Science, Vol. 337, Issue 6096, p. 816-821 https://doi.org/10.1126/science.1225829	journal	June 2012
RNA-Guided Human Genome Engineering via Cas9 Mali, P.; Yang, L.; Esvelt, K. M. Science, Vol. 339, Issue 6121, p. 823-826 https://doi.org/10.1126/science.1232033	journal	January 2013
A double-strand break within a yeast artificial chromosome (YAC) containing human DNA can result in YAC loss, deletion or cell lethality Bennett, C. B.; Westmoreland, T. J.; Snipe, J. R. Molecular and Cellular Biology, Vol. 16, Issue 8 https://doi.org/10.1128/MCB.16.8.4414	journal	August 1996
Multiplex Genome Engineering Using CRISPR/Cas Systems Cong, L.; Ran, F. A.; Cox, D. Science, Vol. 339, Issue 6121, p. 819-823 https://doi.org/10.1126/science.1231143	journal	January 2013
THE CAN1 LOCUS OF SACCHAROMYCES CEREVISIAE: FINE-STRUCTURE ANALYSIS AND FORWARD MUTATION RATES Whelan, William L.; Gocke, Elmar; Manney, Thomas R. Genetics, Vol. 91, Issue 1 https://doi.org/10.1093/genetics/91.1.35	journal	January 1979
Efficient genome editing in zebrafish using a CRISPR-Cas system Hwang, Woong Y.; Fu, Yanfang; Reyon, Deepak Nature Biotechnology, Vol. 31, Issue 3, p. 227-229 https://doi.org/10.1038/nbt.2501	journal	January 2013
Regulatable promoters of Saccharomyces cerevisiae: comparison of transcriptional activity and their use for heterologous expression Mumberg, Dominik; Muller, Rolf; Funk, Martin Nucleic Acids Research, Vol. 22, Issue 25 https://doi.org/10.1093/nar/22.25.5767	journal	January 1994
CRISPR-Cas Systems in Bacteria and Archaea: Versatile Small RNAs for Adaptive Defense and Regulation Bhaya, Devaki; Davison, Michelle; Barrangou, Rodolphe Annual Review of Genetics, Vol. 45, Issue 1, p. 273-297 https://doi.org/10.1146/annurev-genet-110410-132430	journal	December 2011
Ultrafast and memory-efficient alignment of short DNA sequences to the human genome Langmead, Ben; Trapnell, Cole; Pop, Mihai Genome Biology, Vol. 10, Issue 3 https://doi.org/10.1186/gb-2009-10-3-r25	journal	January 2009
RNA-programmed genome editing in human cells Jinek, Martin; East, Alexandra; Cheng, Aaron eLife, Vol. 2, Article No. e00471 https://doi.org/10.7554/eLife.00471	journal	January 2013
A CRISPR Approach to Gene Targeting Carroll, Dana Molecular Therapy, Vol. 20, Issue 9, p. 1658-1660 https://doi.org/10.1038/mt.2012.171	journal	September 2012
Sequence Context Effects on Oligo(dT) Termination Signal Recognition by Saccharomyces cerevisiae RNA Polymerase III Braglia, Priscilla; Percudani, Riccardo; Dieci, Giorgio Journal of Biological Chemistry, Vol. 280, Issue 20 https://doi.org/10.1074/jbc.M412238200	journal	May 2005
Conservative Repair of a Chromosomal Double-Strand Break by Single-Strand DNA through Two Steps of Annealing Storici, Francesca; Snipe, Joyce R.; Chan, Godwin K. Molecular and Cellular Biology, Vol. 26, Issue 20 https://doi.org/10.1128/MCB.00672-06	journal	October 2006
RNA-guided editing of bacterial genomes using CRISPR-Cas systems Jiang, Wenyan; Bikard, David; Cox, David Nature Biotechnology, Vol. 31, Issue 3, p. 233-239 https://doi.org/10.1038/nbt.2508	journal	January 2013
CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III Deltcheva, Elitza; Chylinski, Krzysztof; Sharma, Cynthia M. Nature, Vol. 471, Issue 7340 https://doi.org/10.1038/nature09886	journal	March 2011
High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier Schiestl, Robert H.; Gietz, R. Daniel Current Genetics, Vol. 16, Issue 5-6 https://doi.org/10.1007/bf00340712	journal	December 1989
Genetic and Biochemical Properties of Thialysine-resistant Mutants of Saccharomyces cerevisiae Zwolshen, J. H.; Bhattacharjee, J. K. Microbiology, Vol. 122, Issue 2 https://doi.org/10.1099/00221287-122-2-281	journal	February 1981
The need for speed Flicek, Paul Genome Biology, Vol. 10, Issue 3 https://doi.org/10.1186/gb-2009-10-3-212	journal	January 2009

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