抗原变异可使得多种致病微生物易于逃避宿主免疫应答。表达在感染红细胞表面的恶性疟原虫红细胞表面蛋白1（PfPMP1）与感染红细胞、内皮细胞、树突状细胞以及胎盘的单个或多个受体作用，在黏附及免疫逃避中起关键的作用。每个单倍体基因组var基因家族编码约60种成员，通过启动转录不同的var基因变异体为抗原变异提供了分子基础。

疟原虫var基因转换速率变化导致抗原变异[英]／Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A

Targeted nucleases are powerful tools for mediating genome alteration with high precision. The RNA-guided Cas9 nuclease from the microbial clustered regularly interspaced short palindromic repeats (CRISPR) adaptive immune system can be used to facilitate efficient genome engineering in eukaryotic cells by simply specifying a 20-nt targeting sequence within its guide RNA. Here we describe a set of tools for Cas9-mediated genome editing via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, as well as generation of modified cell lines for downstream functional studies. To minimize off-target cleavage, we further describe a double-nicking strategy using the Cas9 nickase mutant with paired guide RNAs. This protocol provides experimentally derived guidelines for the selection of target sites, evaluation of cleavage efficiency and analysis of off-target activity. Beginning with target design, gene modifications can be achieved within as little as 1-2 weeks, and modified clonal cell lines can be derived within 2-3 weeks.

/pdf/genome-engineering-using-the-crispr-cas9-system-4x8nw03mgu.pdf

Genome engineering using the CRISPR-Cas9 system

The advent of facile genome engineering using the bacterial RNA-guided CRISPR-Cas9 system in animals and plants is transforming biology. We review the history of CRISPR (clustered regularly interspaced palindromic repeat) biology from its initial discovery through the elucidation of the CRISPR-Cas9 enzyme mechanism, which has set the stage for remarkable developments using this technology to modify, regulate, or mark genomic loci in a wide variety of cells and organisms from all three domains of life. These results highlight a new era in which genomic manipulation is no longer a bottleneck to experiments, paving the way toward fundamental discoveries in biology, with applications in all branches of biotechnology, as well as strategies for human therapeutics.

The new frontier of genome engineering with CRISPR-Cas9

http://knockout.cwru.edu/publications/hsucrisprreview.pdf

Development and applications of CRISPR-Cas9 for genome engineering.

https://limlab.ucsf.edu/papers/pdfs/lsq_2013.pdf

Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression.

Organic materials with linearly polarized luminescence (LPL) properties and good processability are critical to the development of advanced optical devices. To fabricate polymer-stabilized thin fil...

Polarized Light Emission from Uniaxially Oriented and Polymer-Stabilized AIE Luminogen Thin Films

Background Adequate fixation of replanted digits is essential not only for short-term healing but for long-term function. Various bony fixation methods using Kirschner (K-) and intraosseous wire are available in replantation. We examined clinical and radiographic outcomes of fixation methods on bone union after digital replantation. Methods A single institutional retrospective review identified 992 patients who had undergone 1,247 successful replantations between July 2009 and September 2015. Exclusion criteria included amputations of the distal phalanx, comminuted fractures, and intra-articular fractures. Patients were classified according to 5 categories of fixation methods: single K-wire, double longitudinal K-wires, cross K-wires, wire with, and wire without K-wire support. Bone union was evaluated by 5-month postoperative X-ray and fixation outcomes were compared across the 5 groups. Results The exclusion criteria were applied, and 88 patients with 103 replanted digits remained for analysis. Single K-wire fixation was used in 40 digits, double longitudinal K-wires in 30, and cross fixation in 14. Wire with and without K-wire support was required in 15 and 4 digits. Nonunion was observed in 32 digits (31.1%), of which 13 required additional operations such as bone graft or corrective osteotomy. The highest percent of nonunion was observed after cross fixation (35.7%) and the lowest after wire alone (25.0%). Conclusions In this study, contrary to general knowledge, we found that single K-wire fixation was not associated with poorer outcomes. Successful bone union outcomes may be achieved by careful selection of bone fixation methods. This study provides useful information for planning bone fixation in digital replantation.

/pdf/analysis-of-bone-fixation-methods-in-digital-replantation-3k1ktwk3wl.pdf

Analysis of Bone Fixation Methods in Digital Replantation

The formation characteristics on the vertical stacks of shape-engineered InAs∕InAlGaAs quantum dots (QDs), which were formed by the alternate growth method (AGQDs), were studied in terms of the modulation in strain field and phase separation. The threshold current of the broad-area laser diodes (LDs) with five stacks of the AGQDs (AGQD-LDs) was 4.5 times smaller than that of the LDs with seven stacks of the conventionally grown QDs (CQD-LDs). The slope efficiency for the AGQD-LDs was 0.16W∕A, which was higher than that of the CQD-LDs of 0.9W∕A. These results can be attributed to better confinement of the electron wave function in QDs.

Vertical stacks of shape-engineered InAs∕InAlGaAs quantum dot and its influences on the lasing characteristics

During the past decade, the density of NAND flash memory has been increased in many folds. The increase has been driven by storing multiple bits in a cell and scaling down the fabrication process. Such advance in manufacturing technology, however, has been significantly impaired the reliability of flash memory so that it becomes one of the major concerns in use of flash memory. Moreover, as flash memory writes data in the unit of flash page, the trend of the increase in page size worsens the reliability by amplifying a small update to a full flash page programming. In this paper, we propose a new programming method to improve the flash endurance cycle, especially when a small amount of data are written repeatedly. The proposed method, so called “subpage programming”, partitions a page into smaller subpages. A small amount of data can be programmed to one of the subpages while the other subpages are inhibited from the programming by leveraging the mechanisms of flash cell programming. Thus, the number of flash cells that undergo programming is minimized. We evaluated the effect of the proposed subpage programming on real NAND flash memory chips from three different manufacturers. Our evaluation results show that subpage programming improves the flash endurance cycle by up to 258%.

/pdf/subpage-programming-for-extending-the-lifetime-of-nand-flash-i8zjyu6wte.pdf

Subpage programming for extending the lifetime of NAND flash memory

The abnormal proliferation of cancer cells is driven by deregulated oncogenes or tumor suppressors, among which the cancer-vulnerable genes are attractive therapeutic targets. Targeting mislocalization of oncogenes and tumor suppressors resulting from aberrant nuclear export is effective for inhibiting growth transformation of cancer cells. We performed a clustered regularly interspaced short palindromic repeat (CRISPR)-associated (Cas) screening in a unique model of matched primary and oncogenic Kaposi's sarcoma-associated herpesvirus (KSHV)-transformed cells and identified genes that were growth promoting and growth suppressive for both types of cells, among which exportin XPO1 was demonstrated to be critical for the survival of transformed cells. Using XPO1 inhibitor KPT-8602 and by small interfering RNA (siRNA) knockdown, we confirmed the essential role of XPO1 in cell proliferation and growth transformation of KSHV-transformed cells and in cell lines of other cancers, including gastric cancer and liver cancer. XPO1 inhibition induced cell cycle arrest through p53 activation, but the mechanisms of p53 activation differed among the different types of cancer cells. p53 activation depended on the formation of promyelocytic leukemia (PML) nuclear bodies in gastric cancer and liver cancer cells. Mechanistically, XPO1 inhibition induced relocalization of autophagy adaptor protein p62 (SQSTM1), recruiting p53 for activation in PML nuclear bodies. Taken the data together, we have identified novel growth-promoting and growth-suppressive genes of primary and cancer cells and have demonstrated that XPO1 is a vulnerable target of cancer cells. XPO1 inhibition induces cell arrest through a novel PML- and p62-dependent mechanism of p53 activation in some types of cancer cells.IMPORTANCE Using a model of oncogenic virus KSHV-driven cellular transformation of primary cells, we have performed a genome-wide CRISPR-Cas9 screening to identify vulnerable genes of cancer cells. This screening is unique in that this virus-induced oncogenesis model does not depend on any cellular genetic alterations and has matched primary and KSHV-transformed cells, which are not available for similar screenings in other types of cancer. We have identified genes that are both growth promoting and growth suppressive in primary and transformed cells, some of which could represent novel proto-oncogenes and tumor suppressors. In particular, we have demonstrated that the exportin XPO1 is a critical factor for the survival of transformed cells. Using a XPO1 inhibitor (KPT-8602) and siRNA-mediated knockdown, we have confirmed the essential role of XPO1 in cell proliferation and in growth transformation of KSHV-transformed cells, as well as of gastric and liver cancer cells. XPO1 inhibition induces cell cycle arrest by activating p53, but the mechanisms of p53 activation differed among different types of cancer cells. p53 activation is dependent on the formation of PML nuclear bodies in gastric and liver cancer cells. Mechanistically, XPO1 inhibition induces relocalization of autophagy adaptor protein p62 (SQSTM1), recruiting p53 for activation in PML nuclear bodies. These results illustrate that XPO1 is a vulnerable target of cancer cells and reveal a novel mechanism for blocking cancer cell proliferation by XPO1 inhibition as well as a novel PML- and p62-mediated mechanism of p53 activation in some types of cancer cells.

/pdf/crispr-cas9-screening-of-kaposi-s-sarcoma-associated-25jmxabaz4.pdf

Jin-Soo Kim

Papers

Polarized Light Emission from Uniaxially Oriented and Polymer-Stabilized AIE Luminogen Thin Films

Analysis of Bone Fixation Methods in Digital Replantation

Vertical stacks of shape-engineered InAs∕InAlGaAs quantum dot and its influences on the lasing characteristics

Subpage programming for extending the lifetime of NAND flash memory

CRISPR-Cas9 Screening of Kaposi's Sarcoma-Associated Herpesvirus-Transformed Cells Identifies XPO1 as a Vulnerable Target of Cancer Cells.