Seasat over-land scatterometer data. II. Selection of extended area and land-target sites for the calibration of spaceborne scatterometers

1989 ◽  
Vol 27 (6) ◽  
pp. 779-788 ◽  
Author(s):  
R.G. Kennett ◽  
F.K. Li
Keyword(s):  
Target Sites ◽  
Extended Area ◽  
Selection Of

scholarly journals The integration preference of Sleeping Beauty at non-TA site is related to the transposon end sequences

2019 ◽  
Author(s):  
Yiting Zhou ◽  
Guangwei Ma ◽  
Jiawen Yang ◽  
Yabin Guo
Keyword(s):  
Site Selection ◽  
Genomic Dna ◽  
Consensus Sequence ◽  
Mouse Cell ◽  
Sleeping Beauty ◽  
Consensus Sequences ◽  
Target Site Selection ◽  
Target Sites ◽  
End Sequences ◽  
Selection Of

Abstract Background: Sleeping Beauty (SB) transposon had been thought to strictly integrate into TA dinucleotides. Recently, we found that SB also integrates into non-TA sites at a lower frequency. Here we performed further study on the non-TA integration of SB. Results: 1) SB can integrate into non-TA sites in HEK293T cells as well as in mouse cell lines. 2) Both the hyperactive transposase SB100X and the traditional SB11 catalyze integrations at non-TA sites. 3) The consensus sequence of the non-TA target sites only occur at the opposite side of the sequenced junction between transposon end and the genomic sequences, indicating that the integrations at non-TA sites are mainly aberrant integrations. 4) The consensus sequence of the non-TA target sites is corresponding to the transposon end sequence. When the transposon end sequence is mutated, the consensus sequences changed too. Conclusion: The interaction between the SB transposon end and genomic DNA may be involved in the target site selection of the SB integrations at non-TA sites.

scholarly journals Establishment of a Screening System for Selection of siRNA Target Sites Directed against Hepatitis B Virus Surface Gene

2005 ◽  
Vol 37 (5) ◽  
pp. 310-316 ◽  
Author(s):  
Xiu-Min ZHOU ◽  
Ju-Sheng LIN ◽  
Yi SHI ◽  
De-An TIAN ◽  
Huan-Jun HUANG ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Screening System ◽  
Virus Surface ◽  
B Virus ◽  
Target Sites ◽  
Sirna Target ◽  
Surface Gene ◽  
Selection Of

scholarly journals PEG-delivered CRISPR-Cas9 Ribonucleoproteins System for Gene-editing Screening of Maize Protoplasts

2020 ◽  
Author(s):  
Rodrigo Ribeiro Arnt Sant’Ana ◽  
Clarissa Alves Caprestano ◽  
Rubens Onofre Nodari ◽  
Sarah Zanon Agapito-Tenfen
Keyword(s):  
Exposure Time ◽  
Dna Sequences ◽  
Mutation Frequency ◽  
Gene Editing ◽  
Inositol Phosphate ◽  
Gc Content ◽  
Assay Method ◽  
Ribonucleoprotein Complexes ◽  
Target Sites ◽  
Selection Of

CRISPR-Cas9 technology allows the modification of DNA sequences in vivo at the location of interest. Although CRISPR-Cas9 can produce genomic changes that do not require DNA vector carriers, the use of transgenesis for stable integration of DNA coding for gene-editing tools into plant genomes is still the most used approach and it can generate unintended transgenic integrations, while Cas9 prolonged expression can increase cleavage at off-target sites. In addition, the selection of genetically modified cells from millions of treated cells, especially plant cells, is still challenging. These downfalls can be avoided with the delivery of preassembled ribonucleoprotein complexes (RNPs) composed of purified recombinant enzyme Cas9 and in vitro- transcribed guide RNA (gRNA) molecules in a protoplast system. We therefore aimed to develop the first DNA-free protocol for gene-editing in maize and introduced RNPs into their protoplasts with PEG 4000. We performed effective transformation of maize protoplasts using different gRNAs sequences targeting the inositol phosphate kinase gene and applying two different exposure times to RNPs. Using low-cost Sanger sequencing protocol, we observed an efficiency rate of 0.85 up to 5.85%, which is equivalent to DNA-free protocols used in other plant species. A positive correlation was displayed between exposure time and mutation frequency. Mutation frequency was gRNA sequence- and exposure time-dependent. In summary, we demonstrated the suitability of RNP transfection as an effective screening platform for gene-editing in maize. This efficient and relatively easy assay method for selection of gRNA suitable for editing of gene of interest will be highly useful for genome editing in maize, since genome size and GC-content are large and high in maize genome, respectively. Nevertheless, the large amplitude of mutations at target site requires scrutiny when checking mutations at off-target sites and potential safety concerns.

scholarly journals The Integration Preference of Sleeping Beauty at Non-TA Site Is Related to the Transposon End Sequences

2021 ◽  
Vol 12 ◽  
Author(s):  
Yiting Zhou ◽  
Guangwei Ma ◽  
Jiawen Yang ◽  
Zenghong Gao ◽  
Yabin Guo
Keyword(s):  
Site Selection ◽  
Genomic Dna ◽  
Consensus Sequence ◽  
Mouse Cell ◽  
Sleeping Beauty ◽  
Consensus Sequences ◽  
Target Site Selection ◽  
Target Sites ◽  
End Sequences ◽  
Selection Of

Recently, we proved that Sleeping Beauty (SB) transposon integrates into non-TA sites at a lower frequency. Here, we performed a further study on the non-TA integration of SB and showed that (1) SB can integrate into non-TA sites in HEK293T cells as well as in mouse cell lines; (2) Both the hyperactive transposase SB100X and the traditional SB11 catalyze integrations at non-TA sites; (3) The consensus sequence of the non-TA target sites only occurs at the opposite side of the sequenced junction between the transposon end and the genomic sequences, indicating that the integrations at non-TA sites are mainly aberrant integrations; and (4) The consensus sequence of the non-TA target sites is corresponding to the transposon end sequence. The consensus sequences changed following the changes of the transposon ends. This result indicated that the interaction between the SB transposon end and genomic DNA (gDNA) may be involved in the target site selection of the SB integrations at non-TA sites.

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