Welcome To Website IAS

Hot news
Achievement

Independence Award

- First Rank - Second Rank - Third Rank

Labour Award

- First Rank - Second Rank -Third Rank

National Award

 - Study on food stuff for animal(2005)

 - Study on rice breeding for export and domestic consumption(2005)

VIFOTEC Award

- Hybrid Maize by Single Cross V2002 (2003)

- Tomato Grafting to Manage Ralstonia Disease(2005)

- Cassava variety KM140(2010)

Centres
Website links
Vietnamese calendar
Library
Visitors summary
 Curently online :  18
 Total visitors :  7453662

High efficiency genome editing using a dmc1 promoter‐controlled CRISPR/Cas9 system in maize
Monday, 2018/04/09 | 07:55:52

Chao Feng, Handong Su , Bai Han , Rui Wang , Yalin Liu , Xianrui Guo , Chang Liu , Jing Zhang , Jing Yuan , James A. Birchler , Fangpu Han

Plant Biotechnology March 23 2018

ABSTRACT

Previous studies revealed that the promoters for driving both Cas9 and sgRNAs are quite important for efficient genome editing by CRISPR/Cas9 in plants. Here we report our results of targeted genome editing using the maize dmc1 gene promoter combined with the U3 promoter for Cas9 and sgRNA, respectively. Three loci in the maize genome were selected for targeting. The T0 plants regenerated were highly‐efficiently edited at the target sites with homozygous or bi‐allelic mutants accounting for about 66%. The mutations in T0 plants could be stably transmitted to the T1 generation and new mutations could be generated in gametes or zygotes. Whole genome re‐sequencing indicated that no off‐target mutations could be detected in the predicted loci with sequence similarity to the targeted site. Our results show that the dmc1 promoter‐controlled (DPC) CRISPR/Cas9 system is highly efficient in maize and provide further evidence that the optimization of the promoters used for the CRISPR/Cas9 system is important for enhancing the efficiency of targeted genome editing in plants. The evolutionary conservation of the dmc1 gene suggests its potential for use in other plant species.

 

See: https://onlinelibrary.wiley.com/doi/abs/10.1111/pbi.12920

Figure. Schematic illustration of DPC CRISPR/Cas9 expression T-DNA. Maize dmc1 gene and U3 promoters were used for Cas9 and sgRNA, respectively.

(a) Schematic illustration of the target site in the zb7 gene. Black box indicates exons, while the lines between them represent introns. Underlined sequence was selected for targeting; ucleotides marked in blue represent PAM (protospacer adjacent motif). (b) RFLP assay of the genomic DNA of 4 transgene-positive calli (#1-#4) in the first batch. For each callus three independent samples were collected and used. Lane 1-3, calli #1; lane 4-6, calli #2; lane 7-9, calli #3; lane 10-12, calli #4; lane13, control (wild type DNA amplicons digested with PvuII). M, DNA marker. Primer pair zb7-F/zb7-R was used for PCR amplification, Pvu II was used for digestion. (c) Mutation analysis of two transgene-positive calli by cloning followed by Sanger sequencing. #3, #4 were the two calli sample with homozygous or bi-allelic mutations by RFLP assay. (d) Sanger sequencing results of mutations in -regenerated seedlings. #1-3, #1-6, #3-8, #4-1 were selected seedlings used for analysis. Left, sequencing chromatograph and right, the edited sequences at the target site. The yellow box indicates the PvuII site.

Back      Print      View: 1507

[ Other News ]___________________________________________________
  • Genome-wide analysis of autophagy-associated genes in foxtail millet (Setaria italica L.) and characterization of the function of SiATG8a in conferring tolerance to nitrogen starvation in rice.
  • Arabidopsis small nucleolar RNA monitors the efficient pre-rRNA processing during ribosome biogenesis
  • XA21-specific induction of stress-related genes following Xanthomonas infection of detached rice leaves.
  • Reducing the Use of Pesticides with Site-Specific Application: The Chemical Control of Rhizoctonia solani as a Case of Study for the Management of Soil-Borne Diseases
  • OsJRL, a rice jacalin-related mannose-binding lectin gene, enhances Escherichia coli viability under high-salinity stress and improves salinity tolerance of rice.
  • Production of lipopeptide biosurfactants by Bacillus atrophaeus 5-2a and their potential use in microbial enhanced oil recovery.
  • GhABF2, a bZIP transcription factor, confers drought and salinity tolerance in cotton (Gossypium hirsutum L.).
  • Resilience of cassava (Manihot esculenta Crantz) to salinity: implications for food security in low-lying regions.
  • Cellulose synthase complexes act in a concerted fashion to synthesize highly aggregated cellulose in secondary cell walls of plants
  • No adverse effects of transgenic maize on population dynamics of endophytic Bacillus subtilis strain B916-gfp
  • Identification and expression analysis of OsLPR family revealed the potential roles of OsLPR3 and 5 in maintaining phosphate homeostasis in rice
  • Functional analysis of molecular interactions in synthetic auxin response circuits
  • Titanium dioxide nanoparticles strongly impact soil microbial function by affecting archaeal nitrifiers.
  • Inducible Expression of the De-Novo Designed Antimicrobial Peptide SP1-1 in Tomato Confers Resistance to Xanthomonas campestris pv. vesicatoria.
  • Toward combined delignification and saccharification of wheat straw by a laccase-containing designer cellulosome
  • SNP-based discovery of salinity-tolerant QTLs in a bi-parental population of rice (Oryza sativa)
  • Pinpointing genes underlying the quantitative trait loci for root-knot nematode resistance in palaeopolyploid soybean by whole genome resequencing.
  • Transcriptome- Assisted Label-Free Quantitative Proteomics Analysis Reveals Novel Insights into Piper nigrum -Phytophthora capsici Phytopathosystem.
  • Brassinosteroids participate in the control of basal and acquired freezing tolerance of plants
  • Rapid hyperosmotic-induced Ca2+ responses in Arabidopsis thaliana exhibit sensory potentiation and involvement of plastidial KEA transporters

 

Designed & Powered by WEBSO CO.,LTD