An archaeal CRISPR type III-B system exhibiting distinctive RNA targeting features and mediating dual RNA and DNA interference

CRISPR-Cas systems provide a small RNA-based mechanism to defend against invasive genetic elements in archaea and bacteria. To investigate the in vivo mechanism of RNA interference by two type III-B systems (Cmr-α and Cmr-β) in Sulfolobus islandicus, a genetic assay was developed using plasmids carrying an artificial mini-CRISPR (AC) locus with a single spacer. After pAC plasmids were introduced into different strains, Northern analyses confirmed that mature crRNAs were produced from the plasmid-borne CRISPR loci, which then guided gene silencing to target gene expression. Spacer mutagenesis identified a trinucleotide sequence in the 3′-region of crRNA that was crucial for RNA interference. Studying mutants lacking Cmr-α or Cmr-β system showed that each Cmr complex exhibited RNA interference. Strikingly, these analyses further revealed that the two Cmr systems displayed distinctive interference features. Whereas Cmr-β complexes targeted transcripts and could be recycled in RNA cleavage, Cmr-α complexes probably targeted nascent RNA transcripts and remained associated with the substrate. Moreover, Cmr-β exhibited much stronger RNA cleavage activity than Cmr-α. Since we previously showed that S. islandicus Cmr-α mediated transcription-dependent DNA interference, the Cmr-α constitutes the first CRISPR system exhibiting dual targeting of RNA and DNA

Heine-Stieltjes correspondence and the polynomial approach to the standard pairing problem

A new approach for solving the Bethe ansatz (Gaudin-Richardson) equations of the standard pairing problem is established based on the Heine-Stieltjes correspondence. For k pairs of valence nucleons on n different single-particle levels, it is found that solutions of the Bethe ansatz equations can be obtained from one (k+1)×(k+1) and one (n-1)×(k+1) matrices, which are associated with the extended Heine-Stieltjes and Van Vleck polynomials, respectively. Since the coefficients in these polynomials are free from divergence with variations in contrast to the original Bethe ansatz equations, the approach provides an efficient and systematic way to solve the problem, which by extension, can also be used to solve a large class of Gaudin-type quantum many-body problems, including an efficient angular momentum projection method for multiparticle systems. © 2012 American Physical Society

The Heine-Stieltjes correspondence and the polynomial approach to the standard pairing problem

A new approach for solving the Bethe ansatz (Gaudin-Richardson) equations of the standard pairing problem is established based on the Heine-Stieltjes correspondence. For $k$ pairs of valence nucleons on $n$ different single-particle levels, it is found that solutions of the Bethe ansatz equations can be obtained from one (k+1)x(k+1) and one (n-1)x(k+1) matrices, which are associated with the extended Heine-Stieltjes and Van Vleck polynomials, respectively. Since the coefficients in these polynomials are free from divergence with variations in contrast to the original Bethe ansatz equations, the approach thus provides with a new efficient and systematic way to solve the problem, which, by extension, can also be used to solve a large class of Gaudin-type quantum many-body problems and to establish a new efficient angular momentum projection method for multi-particle systems.Comment: ReVTeX, 4 pages, no figur

Harnessing type I and type III CRISPR-Cas systems for genome editing

CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR-associated) systems are widespread in archaea and bacteria, and research on their molecular mechanisms has led to the development of genome-editing techniques based on a few Type II systems. However, there has not been any report on harnessing a Type I or Type III system for genome editing. Here, a method was developed to repurpose both CRISPR-Cas systems for genetic manipulation in Sulfolobus islandicus, a thermophilic archaeon. A novel type of genome-editing plasmid (pGE) was constructed, carrying an artificial mini-CRISPR array and a donor DNA containing a non-target sequence. Transformation of a pGE plasmid would yield two alternative fates to transformed cells: wild-type cells are to be targeted for chromosomal DNA degradation, leading to cell death, whereas those carrying the mutant gene would survive the cell killing and selectively retained as transformants. Using this strategy, different types of mutation were generated, including deletion, insertion and point mutations. We envision this method is readily applicable to different bacteria and archaea that carry an active CRISPR-Cas system of DNA interference provided the protospacer adjacent motif (PAM) of an uncharacterized PAM-dependent CRISPR-Cas system can be predicted by bioinformatic analysis

The Capsid Protein of \u3ci\u3eTurnip Crinkle Virus\u3c/i\u3e Overcomes Two Separate Defense Barriers to Facilitate Systemic Movement of the Virus in \u3ci\u3eArabidopsis\u3c/i\u3e

The capsid protein (CP) of Turnip crinkle virus (TCV) is a multifunctional protein needed for virus assembly, suppression of RNA silencing-based antiviral defense, and long-distance movement in infected plants. In this report, we have examined genetic requirements for the different functions of TCV CP and evaluated the interdependence of these functions. A series of TCV mutants containing alterations in the CP coding region were generated. These alterations range from single-amino-acid substitutions and domain truncations to knockouts of CP translation. The latter category also contained two constructs in which the CP coding region was replaced by either the cDNA of a silencing suppressor of a different virus or that of green fluorescent protein. These mutants were used to infect Arabidopsis plants with diminished antiviral silencing capability (dcl2 dcl3 dcl4 plants). There was a strong correlation between the ability of mutants to reach systemic leaves and the silencing suppressor activity of mutant CP. Virus particles were not essential for entry of the viral genome into vascular bundles in the inoculated leaves in the absence of antiviral silencing. However, virus particles were necessary for egress of the viral genome from the vasculature of systemic leaves. Our experiments demonstrate that TCV CP not only allows the viral genome to access the systemic movement channel through silencing suppression but also ensures its smooth egress by way of assembled virus particles. These results illustrate that efficient long-distance movement of TCV requires both functions afforded by the CP

The Capsid Protein of \u3ci\u3eTurnip Crinkle Virus\u3c/i\u3e Overcomes Two Separate Defense Barriers to Facilitate Systemic Movement of the Virus in \u3ci\u3eArabidopsis\u3c/i\u3e

The capsid protein (CP) of Turnip crinkle virus (TCV) is a multifunctional protein needed for virus assembly, suppression of RNA silencing-based antiviral defense, and long-distance movement in infected plants. In this report, we have examined genetic requirements for the different functions of TCV CP and evaluated the interdependence of these functions. A series of TCV mutants containing alterations in the CP coding region were generated. These alterations range from single-amino-acid substitutions and domain truncations to knockouts of CP translation. The latter category also contained two constructs in which the CP coding region was replaced by either the cDNA of a silencing suppressor of a different virus or that of green fluorescent protein. These mutants were used to infect Arabidopsis plants with diminished antiviral silencing capability (dcl2 dcl3 dcl4 plants). There was a strong correlation between the ability of mutants to reach systemic leaves and the silencing suppressor activity of mutant CP. Virus particles were not essential for entry of the viral genome into vascular bundles in the inoculated leaves in the absence of antiviral silencing. However, virus particles were necessary for egress of the viral genome from the vasculature of systemic leaves. Our experiments demonstrate that TCV CP not only allows the viral genome to access the systemic movement channel through silencing suppression but also ensures its smooth egress by way of assembled virus particles. These results illustrate that efficient long-distance movement of TCV requires both functions afforded by the CP

Colloidal quantum dot hybrids: an emerging class of materials for ambient lighting

The rapid growth of the global economy and urbanization have resulted in major worldwide issues such as greenhouse gas emission, air pollution and the energy crisis. Artificial ambient light is one of the greatest inventions in human history, but it is also one of the primary energy consumption constituents and a focus of the global grand energy challenge. Therefore, low cost and low energy consumption lighting technology is in high demand. In this review, we will summarise and discuss one of the emerging lighting technologies – white electroluminescence light-emitting diodes enabled by hybrid colloidal quantum dots (WQLEDs), which have attracted intense attention because of promising potential in both flat-panel backlighting and solid-state lighting. WQLEDs have unique high luminescence efficiency, broad colour tunability and solution processability. Over the past few decades, the development of colloidal quantum dot synthesis, material engineering and device architecture has highlighted the tremendous improvements in WQLED formation. As WQLED efficiencies approach those of molecular organic LEDs, we identify the critical scientific and technological challenges and provide an outlook for ongoing strategies to overcome these challenges