scholarly journals 310. High-Throughput Gene Editing in Human Cells Enabled by Fast Ligation-Based Automatable Solid-Phase High-Throughput (FLASH) Assembly of TALENs

2012 ◽  
Vol 20 ◽  
pp. S122-S123
Keyword(s):  
High Throughput ◽  
Gene Editing ◽  
Solid Phase ◽  
Human Cells

scholarly journals High-Throughput Profiling of Cas12a Orthologues and Engineered Variants for Enhanced Genome Editing Activity

2021 ◽  
Vol 22 (24) ◽  
pp. 13301
Author(s):  
Dan Zhu ◽  
Junyi Wang ◽  
Di Yang ◽  
Jianzhong Xi ◽  
Juan Li
Keyword(s):  
High Throughput ◽  
Gene Editing ◽  
Genome Engineering ◽  
Human Cells ◽  
Wild Type ◽  
Guide Rnas ◽  
Promising Tool ◽  
Target Sites ◽  
Editing Activity ◽  
Type V

CRISPR/Cas12a (formerly Cpf1), an RNA-guided endonuclease of the Class II Type V-A CRISPR system, provides a promising tool for genome engineering. Over 10 Cas12a orthologues have been identified and employed for gene editing in human cells. However, the functional diversity among emerging Cas12a orthologues remains poorly explored. Here, we report a high-throughput comparative profiling of editing activities across 16 Cas12a orthologues in human cells by constructing genome-integrated, self-cleaving, paired crRNA–target libraries containing >40,000 guide RNAs. Three Cas12a candidates exhibited promising potential owing to their compact structures and editing efficiency comparable with those of AsCas12a and LbCas12a, which are well characterized. We generated three arginine substitution variants (3Rv) via structure-guided protein engineering: BsCas12a-3Rv (K155R/N512R/K518R), PrCas12a-3Rv (E162R/N519R/K525R), and Mb3Cas12a-3Rv (D180R/N581R/K587R). All three Cas12a variants showed enhanced editing activities and expanded targeting ranges (NTTV, NTCV, and TRTV) compared with the wild-type Cas12a effectors. The base preference analysis among the three Cas12a variants revealed that PrCas12a-3Rv shows the highest activity at target sites with canonical PAM TTTV and non-canonical PAM TTCV, while Mb3Cas12a-3Rv exhibits recognition features distinct from the others by accommodating for more nucleotide A at position −3 for PAM TATV and at position −4 for PAM ATCV. Thus, the expanded Cas12a toolbox and an improved understanding of Cas12a activities should facilitate their use in genome engineering.

High-throughput solid-phase permethylation of glycans prior to mass spectrometry

2008 ◽  
Vol 22 (5) ◽  
pp. 721-734 ◽  
Author(s):  
Pilsoo Kang ◽  
Yehia Mechref ◽  
Milos V. Novotny
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
Solid Phase

A high-throughput nanofibers mat-based micro-solid phase extraction for the determination of cationic dyes in wastewater

2016 ◽  
Vol 1460 ◽  
pp. 24-32 ◽  
Author(s):  
Feifei Qi ◽  
Liangliang Qian ◽  
Jingjing Liu ◽  
Xiaoqing Li ◽  
Lingeng Lu ◽  
...  
Keyword(s):  
Solid Phase Extraction ◽  
High Throughput ◽  
Solid Phase ◽  
Cationic Dyes ◽  
Phase Extraction

Analysis of Microbial Community Structure and Volatile Compounds in the Pit Mud Used for Manufacturing Taorong-type Baijiu Based on High-Throughput Sequencing

2021 ◽  
Author(s):  
Yanbo Liu ◽  
Mengxiao Sun ◽  
Pei Hou ◽  
Wenya Wang ◽  
Xiangkun Shen ◽  
...  
Keyword(s):  
Community Structure ◽  
Microbial Community ◽  
Qualitative Analysis ◽  
Volatile Compounds ◽  
High Throughput ◽  
Microbial Community Structure ◽  
High Throughput Sequencing ◽  
Solid Phase ◽  
Gas Chromatography Mass Spectrometry ◽  
Pit Mud

Abstract In this study, the pit mud used in manufacturing Taorong-type Baijiu was collected from the upper, middle, lower and bottom layers of pits in Henan Yangshao Liquor Co., LTD. Besides, high-throughput sequencing (HTS) technology was adopted to analyze the microbial community structure of the pit mud. In addition, the volatile compounds in the pit mud were subjected to preliminarily qualitative analysis through headspace-solid phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS). The results of HTS demonstrated that there were 5, 3, 5 and 5 dominant bacterial phyla (including 11, 11, 9 and 8 dominant bacterial genera) and 3, 3, 3 and 3 dominant fungal phyla (including 4, 7, 7 and 5 dominant fungal genera) in the pit mud from F-S (upper), G-Z (middle), H-X (lower) and I-D (bottom), respectively. The qualitative analysis results of volatile compounds demonstrated that a total of 78 kinds of volatile compounds were detected in the pit mud, including 46, 45, 39 and 49 kinds in the pit mud from F-S, G-Z, H-X and I-D, respectively. Ester and acid were the two main components in the pit mud. Meanwhile, the correlation between microorganisms and main volatile compounds in the pit mud was analyzed. Moreover, Lentimicrobium, Syner-01 and Blvii28_wastewater-sludge group were found for the first time in the pit mud used for manufacturing Taorong-type Baijiu. The findings of this study could provide a theoretical foundation for improving the quality of pit mud and the flavor of Taorong-type Baijiu.

scholarly journals A High-Throughput Search for Composition–Phase–Property Relations of Cu–Ni/Ti–Al Elastic Copper Alloys

Materials ◽  
2018 ◽  
Vol 11 (12) ◽  
pp. 2513
Author(s):  
Wang Jianwei ◽  
Xiao Wei ◽  
Zhang Chuan ◽  
Sun Lu ◽  
Huang Guojie ◽  
...  
Keyword(s):  
High Throughput ◽  
Solid Phase ◽  
Copper Alloys ◽  
Ternary Systems ◽  
Diagram Method ◽  
Property Relations ◽  
Diffusion Layers ◽  
Heat Treated ◽  
Phase Property ◽  
Relationship Of

A high-throughput method was employed to effectively obtain the cross-scale relationship of elastic copper alloys. Firstly, a Cu–Ni–Ti–Cu25Al–Cu35Sn diffusion multiple was prepared and heat-treated under a specified condition to form a series of diffusion layers with the concentration gradient at the multielement metal interface. Then, the compositions, elastic moduli, and hardness of the Cu–Ni–Al and Cu–Ti–Al ternary copper alloys were tested. Meanwhile, the solid phase sequences in the diffusion zones were predicted by the CALPHAD (CALculation of PHAse Diagram) method. Through these experimental and calculated results, the composition–phase–property relations of the Cu–Ni–Al and Cu–Ti–Al ternary systems were established.

scholarly journals High-Throughput Analysis of Selected Urinary Hydroxy Polycyclic Aromatic Hydrocarbons by an Innovative Automated Solid-Phase Microextraction

Molecules ◽  
2018 ◽  
Vol 23 (8) ◽  
pp. 1869 ◽  
Author(s):  
Stefano Dugheri ◽  
Alessandro Bonari ◽  
Matteo Gentili ◽  
Giovanni Cappelli ◽  
Ilenia Pompilio ◽  
...  
Keyword(s):  
Polycyclic Aromatic Hydrocarbons ◽  
High Throughput ◽  
Aromatic Hydrocarbons ◽  
High Throughput Screening ◽  
Solid Phase Microextraction ◽  
Solid Phase ◽  
User Friendliness ◽  
High Throughput Analysis ◽  
Polycyclic Aromatic ◽  
User Friendly

High-throughput screening of samples is the strategy of choice to detect occupational exposure biomarkers, yet it requires a user-friendly apparatus that gives relatively prompt results while ensuring high degrees of selectivity, precision, accuracy and automation, particularly in the preparation process. Miniaturization has attracted much attention in analytical chemistry and has driven solvent and sample savings as easier automation, the latter thanks to the introduction on the market of the three axis autosampler. In light of the above, this contribution describes a novel user-friendly solid-phase microextraction (SPME) off- and on-line platform coupled with gas chromatography and triple quadrupole-mass spectrometry to determine urinary metabolites of polycyclic aromatic hydrocarbons 1- and 2-hydroxy-naphthalene, 9-hydroxy-phenanthrene, 1-hydroxy-pyrene, 3- and 9-hydroxy-benzoantracene, and 3-hydroxy-benzo[a]pyrene. In this new procedure, chromatography’s sensitivity is combined with the user-friendliness of N-tert-butyldimethylsilyl-N-methyltrifluoroacetamide on-fiber SPME derivatization using direct immersion sampling; moreover, specific isotope-labelled internal standards provide quantitative accuracy. The detection limits for the seven OH-PAHs ranged from 0.25 to 4.52 ng/L. Intra-(from 2.5 to 3.0%) and inter-session (from 2.4 to 3.9%) repeatability was also evaluated. This method serves to identify suitable risk-control strategies for occupational hygiene conservation programs.

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