scholarly journals Efficient, selectable marker free gene targeting in soybean using novel Ochrobactrum haywardense-mediated delivery

2021 ◽  
Author(s):  
Sandeep Kumar ◽  
Zhan-Bin Lui ◽  
Nathalie Sanyour-Doyel ◽  
Brian Landerts ◽  
Andrew Worden ◽  
...  
Keyword(s):  
High Resolution ◽  
Gene Targeting ◽  
Plasmid Dna ◽  
Selectable Marker ◽  
Low Frequency ◽  
Transformation Method ◽  
Dna Templates ◽  
Marker Free ◽  
Free Gene ◽  
Donor Template

We report robust selectable marker-free gene targeting (GT) system in soybean, one of the most economically important crops. A novel efficient Ochrobactrum haywardense-mediated embryonic axis transformation method was used for the delivery of CRISPR-Cas9 components and donor template to regenerate T0 plants in 6-8 weeks after transformation. This approach generated up to 3.4% targeted insertion of the donor sequence into the target locus in T0 plants, with ~ 90% mutation rate observed at the genomic target site. The GT was demonstrated in two genomic sites using two different donor DNA templates without a need of a selectable marker within the template. High-resolution Southern by Sequencing (SbS) analysis identified T1 plants with precise targeted insertion and without unintended plasmid DNA. Unlike previous low-frequency GT reports in soybean that involved particle bombardment-mediated delivery and extensive selection, the method described here is fast, efficient, reproducible, does not require selectable marker within the donor DNA, and generates non-chimeric plants with heritable GT.

scholarly journals Efficient Gene Targeting in Maize using Inducible CRISPR-Cas9 and Marker-Free Donor Template

2020 ◽  
Author(s):  
Pierluigi Barone ◽  
Emily Wu ◽  
Brian Lenderts ◽  
Ajith Anand ◽  
William Gordon-Kamm ◽  
...  
Keyword(s):  
Gene Targeting ◽  
Selectable Marker ◽  
Marker Gene ◽  
In Planta ◽  
Genomic Deletions ◽  
Efficient Gene ◽  
Marker Free ◽  
Genomic Gene ◽  
Donor Locus ◽  
Donor Template

AbstractCRISPR-Cas9 is a powerful tool for generating targeted mutations and genomic deletions. However, precise gene insertion or sequence replacement remains a major hurdle before application of CRISPR-Cas9 technology is fully realized in plant breeding. Here we report high frequency, selectable marker-free intra-genomic gene targeting (GT) in maize. Heat shock-inducible Cas9 was used for generating targeted double-strand breaks (DSBs) and simultaneous mobilization of the donor template from pre-integrated T-DNA. The construct was designed such that release of the donor template and subsequent DNA repair activated expression of the selectable marker gene within the donor locus. This approach generated up to 4.7% targeted insertion of the donor sequence into the target locus in T0 plants, with up to 86% detected donor template release and 99% mutation rate were observed at the donor loci and the genomic target site, respectively. Unlike previous in planta or intra-genomic homologous recombination reports, that required multiple generations and extensive screening, our method provides non-chimeric, heritable GT in the T0 generation.

Marker-Free Gene Targeting by Recombinase-Mediated Cassette Exchange

2012 ◽  
pp. 379-390 ◽  
Author(s):  
Hiroyasu Ebinuma ◽  
Kazuya Nanto ◽  
Saori Kasahara ◽  
Atsushi Komamine
Keyword(s):  
Gene Targeting ◽  
Cassette Exchange ◽  
Marker Free ◽  
Free Gene

scholarly journals Efficient Gene Targeting in Maize Using Inducible CRISPR-Cas9 and Marker-free Donor Template

2020 ◽  
Vol 13 (8) ◽  
pp. 1219-1227 ◽  
Author(s):  
Pierluigi Barone ◽  
Emily Wu ◽  
Brian Lenderts ◽  
Ajith Anand ◽  
William Gordon-Kamm ◽  
...  
Keyword(s):  
Gene Targeting ◽  
Efficient Gene ◽  
Marker Free ◽  
Donor Template

Practical High Resolution Microscopy

1968 ◽  
Vol 26 ◽  
pp. 302-303
Author(s):  
P. A. Marsh ◽  
T. Mullens ◽  
D. Price
Keyword(s):  
High Resolution ◽  
Magnetic Fields ◽  
Low Frequency ◽  
Resolving Power ◽  
Careful Attention ◽  
Electron Microscopes ◽  
The Relationship ◽  
High Resolution Microscopy ◽  
New Facilities

It is possible to exceed the guaranteed resolution on most electron microscopes by careful attention to microscope parameters essential for high resolution work. While our experience is related to a Philips EM-200, we hope that some of these comments will apply to all electron microscopes.The first considerations are vibration and magnetic fields. These are usually measured at the pre-installation survey and must be within specifications. It has been our experience, however, that these factors can be greatly influenced by the new facilities and therefore must be rechecked after the installation is completed. The relationship between the resolving power of an EM-200 and the maximum tolerable low frequency interference fields in milli-Oerstedt is 10 Å - 1.9, 8 Å - 1.4, 6 Å - 0.8.

Rapid visualized assessment of drug efficacy in live mice with a selectable marker-free autoluminescent Klebsiella pneumoniae

2021 ◽  
Vol 177 ◽  
pp. 112919
Author(s):  
Xirong Tian ◽  
Yamin Gao ◽  
Shuai Wang ◽  
H.M. Adnan Hameed ◽  
Wei Yu ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Selectable Marker ◽  
Drug Efficacy ◽  
Marker Free

Nanoscale structures of circle - MgCl2 constructed by plasmid DNA templates

2005 ◽  
Vol 37 (3) ◽  
pp. 151-161 ◽  
Author(s):  
Aiguo Wu ◽  
Zhuang Li ◽  
Hualan Zhou ◽  
Erkang Wang
Keyword(s):  
Plasmid Dna ◽  
Dna Templates ◽  
Nanoscale Structures

scholarly journals Low frequency (74 MHz) radio continuum observations of the inner 13° × 7° of the Galactic center

2013 ◽  
Vol 9 (S303) ◽  
pp. 464-466
Author(s):  
M. Rickert ◽  
F. Yusef-Zadeh ◽  
C. Brogan
Keyword(s):  
High Resolution ◽  
Molecular Cloud ◽  
Supernova Remnant ◽  
Galactic Center ◽  
Low Frequency ◽  
Radio Continuum ◽  
Large Array ◽  
Very Large Array ◽  
Image Displays ◽  
Western Side

AbstractWe analyze a high resolution (114″ × 60″) 74 MHz image of the Galactic center taken with the Very Large Array (VLA). We have identified several absorption and emission features in this region, and we discuss preliminary results of two Galactic center sources: the Sgr D complex (G1.1–0.1) and the Galactic center lobe (GCL).The 74 MHz image displays the thermal and nonthermal components of Sgr D and we argue the Sgr D supernova remnant (SNR) is consistent with an interaction with a nearby molecular cloud and the location of the Sgr D Hii region on the near side of the Galactic center. The image also suggests that the emission from the eastern side of the GCL contains a mixture of both thermal and nonthermal sources, whereas the western side is primarily thermal.

High-resolution and wideband optical vector analysis using fixed low-frequency detection

2021 ◽  
Author(s):  
Ting Qing ◽  
Shupeng Li ◽  
Lihan Wang ◽  
Xiaohu Tang ◽  
Ping Li ◽  
...  
Keyword(s):  
High Resolution ◽  
Low Frequency ◽  
Vector Analysis ◽  
Frequency Detection

COMPARATIVE ANALYSIS OF SPECTRA OF THE 2000-YEAR NORTHERN HEMISPHERE AVERAGE SURFACE AIR TEMPERATURE RECONSTRUCTIONS

2021 ◽  
pp. 5-13
Author(s):  
N. M. DATSENKO ◽  
◽  
D. M. SONECHKIN ◽  
B. YANG ◽  
J.-J. LIU ◽  
...  
Keyword(s):  
High Resolution ◽  
Northern Hemisphere ◽  
Air Temperature ◽  
Wavelet Transforms ◽  
Spectral Composition ◽  
Low Frequency ◽  
Surface Air Temperature ◽  
Temporal Variations ◽  
Continuous Wavelet Transforms ◽  
Stable Estimation

The spectral composition of temporal variations in the Northern Hemisphere mean surface air temperature is estimated and compared in 2000-year paleoclimatic reconstructions. Continuous wavelet transforms of these reconstructions are used for the stable estimation of energy spectra. It is found that low-frequency parts of the spectra (the periods of temperature variations of more than 100 years) based on such high-resolution paleoclimatic indicators as tree rings, corals, etc., are similar to the spectrum of white noise, that is never observed in nature. This seems unrealistic. The famous reconstruction called “Hockey Stick” is among such unrealistic reconstructions. Reconstructions based not only on high-resolution but also on low-resolution indicators seem to be more realistic, since the low-frequency parts of their spectra have the pattern of red noise. They include the “Boomerang” reconstruction showing that some warm periods close to the present-day one were observed in the past.

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