Efficient gene replacement by CRISPR/Cas-mediated homologous recombination in the model diatom Thalassiosira pseudonana

CRISPR/Cas enables targeted genome editing in many different plant and algal species including the model diatom Thalassiosira pseudonana. However, efficient gene targeting by homologous recombination (HR) to date is only reported for photosynthetic organisms in their haploid life-cycle phase. Here, a CRISPR/Cas construct, assembled using Golden Gate cloning, enabled highly efficient HR in a diploid photosynthetic organism. Homologous recombination was induced in T. pseudonana using sequence-specific CRISPR/Cas, paired with a dsDNA donor matrix, generating substitution of the silacidin, nitrate reductase and urease genes by a resistance cassette (FCP:NAT). Up to c. 85% of NAT-resistant T. pseudonana colonies screened positive for HR by nested PCR. Precise integration of FCP:NAT at each locus was confirmed using an inverse PCR approach. The knockout of the nitrate reductase and urease genes impacted growth on nitrate and urea, respectively, while the knockout of the silacidin gene in T. pseudonana caused a significant increase in cell size, confirming the role of this gene for cell-size regulation in centric diatoms. Highly efficient gene targeting by HR makes T. pseudonana as genetically tractable as Nannochloropsis and Physcomitrella, hence rapidly advancing functional diatom biology, bionanotechnology and biotechnological applications targeted on harnessing the metabolic potential of diatoms

Local Hamiltonians Whose Ground States Are Hard to Approximate

Ground states of local Hamiltonians can be generally highly entangled: Any quantum circuit that generates them (even approximately) must be sufficiently deep to allow coupling (entanglement) between any pair of qubits. Until now this property was not known to be robust - the marginals of such states to a subset of the qubits containing all but a small constant fraction of them may be only locally entangled, and hence approximable by shallow quantum circuits. In this work we construct a family of 16-local Hamiltonians for which any 1-10[superscript-8] fraction of qubits of any ground state must be highly entangled.This provides evidence that quantum entanglement is not very fragile, and perhaps our intuition about its instability is an artifact of considering local Hamiltonians which are not only local but spatially local. Formally, it provides positive evidence for two wide-open conjectures in condensed-matter physics and quantum complexity theory which are the qLDPC conjecture, positing the existence of good quantum LDPC codes, and the NLTS conjecture due to Freedman and Hastings positing the existence of local Hamiltonians in which any low-energy state is highly-entangled.Our Hamiltonian is based on applying the hypergraph product by Tillich-Zemor to the repetition code with checks from an expander graph. A key tool in our proof is a new lower bound on the vertex expansion of the output of low-depth quantum circuits, which may be of independent interest. Keywords: quantum entanglement; stationary state; robustness; complexity theory; graph theoryNational Science Foundation (U.S.) (Grant CCF-1111382)National Science Foundation (U.S.) (Grant CCF-1452616)National Science Foundation (U.S.) (Grant CCF-1629809)United States. Army Research Office (Contract W911NF-12-1-0486