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.

Factorial Stirling matrix and related combinatorial sequences

Enrichment of cells with TALEN-induced mutations using surrogate reporters.

/pdf/response-to-unexpected-mutations-after-crispr-cas9-editing-ougzch24zw.pdf

Response to "unexpected mutations after CRISPR-Cas9 editing in vivo"

In this paper, low-resistivity molybdenum (Mo) thin films were deposited on stainless-steel and soda-lime glass substrates with good adhesion at room temperature by using a direct current (DC) magnetron technique. The Mo thin films were annealed at different temperatures in order to enhance their structural and electrical properties so that they could be used for the solar cell applications. The Mo thin film is the bottom electrode used in the chalcopyrite solar cell family (CuInSe{sub 2} and its alloys) because of the low resistivity of the Cu(InGa)Se{sub 2}/Mo contact. X-ray diffraction (XRD) analysis showed that these Mo films were polycrystalline in nature and exhibited better crystallization with increasing crystallite size as the annealing temperature was raised from 400 to 800 .deg. C. The Mo film's crystallite size was observed to increase from 80 to 450 nm as the annealing temperature was increased from as-deposited to 800 .deg. C. The microstructure, surface morphology and homogenous grains, were observed using field emission scanning electron microscopy (FE-SEM) and atomic force microscopy (AFM).

Jin-Soo Kim

Papers

Factorial Stirling matrix and related combinatorial sequences

Enrichment of cells with TALEN-induced mutations using surrogate reporters.

Response to "unexpected mutations after CRISPR-Cas9 editing in vivo"

Effect of Thermal Annealing on the Structure, Morphology, and Electrical Properties of Mo Bottom Electrodes for Solar Cell Applications

Peptide tags for a dual affinity fusion system.