A Conformational Restriction Strategy for the Control of CRISPR/Cas Gene Editing with Photoactivatable Guide RNAs.

The CRISPR/Cas system is one of the most powerful tools for gene editing. However, approaches for precise control of genome editing and regulatory events are still desirable. Here, we report the spatiotemporal and efficient control of CRISPR/Cas9- and Cas12a-mediated editing with conformationally restricted guide RNAs (gRNAs). This approach relied on only two or three pre-installed photo-labile substituents followed by an intramolecular cyclization, representing a robust synthetic method in comparison to the heavily modified linear gRNAs that often require extensive screening and time-consuming optimization. This tactic could direct the precise cleavage of the genes encoding green fluorescent protein (GFP) and the vascular endothelial growth factor A (VEGFA) protein within a predefined cutting region without notable editing leakage in live cells. We also achieved light-mediated myostatin (MSTN) gene editing in embryos, wherein a new bow-knot-type gRNA was constructed with excellent OFF/ON switch efficiency. Overall, our work provides a significant new strategy in CRISPR/Cas editing with modified circular gRNAs to precisely manipulate where and when genes are edited.

Metropolitan all-pass and inter-city quantum communication network.

We have demonstrated a metropolitan all-pass quantum communication network in field fiber for four nodes. Any two nodes of them can be connected in the network to perform quantum key distribution (QKD). An optical switching module is presented that enables arbitrary 2-connectivity among output ports. Integrated QKD terminals are worked out, which can operate either as a transmitter, a receiver, or even both at the same time. Furthermore, an additional link in another city of 60 km fiber (up to 130 km) is seamless integrated into this network based on a trusted relay architecture. On all the links, we have implemented protocol of decoy state scheme. All of necessary electrical hardware, synchronization, feedback control, network software, execution of QKD protocols are made by tailored designing, which allow a completely automatical and stable running. Our system has been put into operation in Hefei in August 2009, and publicly demonstrated during an evaluation conference on quantum network organized by the Chinese Academy of Sciences on August 29, 2009. Real-time voice telephone with one-time pad encoding between any two of the five nodes (four all-pass nodes plus one additional node through relay) is successfully established in the network within 60 km.

Anti-Ki-67 peptide nucleic acid affects the proliferation and apoptosis of human renal carcinoma cells in vitro.

We treated in vitro human renal carcinoma cells (cell line 786-0) with the lipid-delivered peptide nucleic acids (PNAs) against Ki-67 gene. Corresponding control groups were treated with the antisense oligonucleotides (ASOs) of the same nucleobase sequence, and with mismatched PNAs. In cells treated by anti-Ki-67 PNAs, the Ki-67 expression rate, Ki-67 protein level, cell growth and the DNA synthesis-indicative 3H-thymidine incorporation rate were lower than in the ASO-treated groups, and reduced significantly compared to untreated controls, whereas the rate of apoptosis was markedly increased by PNA treatment. We conclude that anti-Ki-67 PNA has more strong (than ASO) and dose-dependent effects on the proliferation and apoptosis of human renal carcinoma cells. Our results indicate that the strategy of using PNA against the Ki-67 gene might be a promising approach in renal carcinoma therapy.