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.

Renal impairment (RI) is a relatively common complication of multiple myeloma, which increases in frequency as disease becomes more advanced and recovery of renal function becomes less likely as patients progress through lines of therapy. Clinical trials in the relapsed/refractory multiple myeloma (RRMM) setting have not uniformly included patients with RI or robustly reported their outcomes. Here, we review existing data among patients with RI and RRMM across drug classes (including immunomodulatory agents, proteasome inhibitors, monoclonal antibodies, antibody-drug conjugates, chimeric antigen receptor T-cell therapies, and exportin-1 inhibitor) to provide an improved understanding of available treatment options for this important population. We highlight data from pivotal clinical trials, including data relating to renal response (as defined by the International Myeloma Working Group) and discuss real-world experiences in patients with RI, where applicable. Despite substantial advances in RRMM treatment, the presence of RI remains associated with reduced overall survival. Consistent inclusion of patients with RI, and uniform reporting of their outcomes, should be encouraged in future prospective trials of treatments for RRMM.

An overview of treatment options for patients with relapsed/refractory multiple myeloma and renal impairment

Although several high-fidelity SpCas9 variants have been reported, it has been observed that this increased specificity is associated with reduced on-target activity, limiting the applications of the high-fidelity variants when efficient genome editing is required. Here, we developed an improved version of Sniper–Cas9, Sniper2L, which represents an exception to this trade-off trend as it showed higher specificity with retained high activity. We evaluated Sniper2L activities at a large number of target sequences and developed DeepSniper, a deep learning model that can predict the activity of Sniper2L. We also confirmed that Sniper2L can induce highly efficient and specific editing at a large number of target sequences when it is delivered as a ribonucleoprotein complex. Mechanically, the high specificity of Sniper2L originates from its superior ability to avoid unwinding a target DNA containing even a single mismatch. We envision that Sniper2L will be useful when efficient and specific genome editing is required. Kim et al. used directed evolution methods to identify a high-fidelity SpCas9 variant, Sniper2L, which exhibits high general activity but maintains high specificity at a large number of target sites.

Sniper2L is a high-fidelity Cas9 variant with high activity

【課題】標的ＤＮＡに特異的なガイドＲＮＡおよびＣａｓタンパク質コード核酸もしくはＣａｓタンパク質を含む、真核細胞または真核生物中の標的ＤＮＡを切断するための組成物の提供。 【解決手段】真核細胞における標的ＤＮＡを改変する方法であって、ＣＲＩＳＰＲ ＲＮＡ（ｃｒＲＮＡ）およびトランス活性化ｃｒＲＮＡ（ｔｒａｃｒＲＮＡ）を含むキメラＲＮＡであるガイドＲＮＡを調製し、ここで標的ＤＮＡは、ガイドＲＮＡのｃｒＲＮＡに相補的な領域を有する第１の鎖、及びプロトスペーサー隣接モチーフ（ＰＡＭ）を有する第２の鎖を含み、第２の鎖上のＰＡＭを認識するＣａｓタンパク質を調製し、ガイドＲＮＡをＣａｓタンパク質とｉｎ ｖｉｔｒｏで複合体形成させ、標的ＤＮＡを有する真核細胞にＣａｓタンパク質/ガイドＲＮＡ複合体を移入させる、ことを含む、上記方法。 【選択図】図１ａ

標的ｄｎａに特異的なガイドｒｎａおよびｃａｓタンパク質コード核酸またはｃａｓタンパク質を含む、標的ｄｎａを切断するための組成物、ならびにその使用

Effect of High Dose Oral Rabeprazole On Recurrent Bleeding After Endoscopic Treatment of Bleeding Peptic Ulcers: A Preliminary Report

Recently, molybdenum diselenide (MoSe2) has attracted nascent research attention for potential applications in electronic and optoelectronic devices due to its unique properties including tunable bandgap, strong photoluminescence and large exciton binding energy. However,\n the synthesis of reproducible, controlled and large scale MoSe2 films is still a great challenge. Here, we have investigated the morphology, structure and crystalline quality of MoSe2 films synthesized by the selenization of Mo metal films. The Mo metal films of different\n thicknesses were deposited at room temperature by direct current sputtering. Subsequently, MoSe2 films were prepared by selenization of sputtered Mo films at 550 °C for 20 minutes. The obtained MoSe2 films are polycrystalline with hexagonal crystal structure. The\n crystalline quality of the MoSe2 films is improved with increase in the thickness of Mo metal films. The MoSe2 films are found to be n-type in nature and reasonably stoichiometric (Mo/Se ratio ∼1:1.9). This study provides an experimental demonstration of an\n alternative cost-effective direct synthesis of MoSe2 films on SiO2/Si for the applications of semiconductor devices.

Jin-Soo Kim

Papers

An overview of treatment options for patients with relapsed/refractory multiple myeloma and renal impairment

Sniper2L is a high-fidelity Cas9 variant with high activity

標的ｄｎａに特異的なガイドｒｎａおよびｃａｓタンパク質コード核酸またはｃａｓタンパク質を含む、標的ｄｎａを切断するための組成物、ならびにその使用

Effect of High Dose Oral Rabeprazole On Recurrent Bleeding After Endoscopic Treatment of Bleeding Peptic Ulcers: A Preliminary Report

Direct Synthesis of MoSe2 Thin Films on SiO2/Si Using Selenization Process of Sputtered Molybdenum