MRE11 and RAD50, but not NBS1, are essential for gene targeting in the moss Physcomitrella patens

The moss Physcomitrella patens is unique among plant models for the high frequency with which targeted transgene insertion occurs via homologous recombination. Transgene integration is believed to utilize existing machinery for the detection and repair of DNA double-strand breaks (DSBs). We undertook targeted knockout of the Physcomitrella genes encoding components of the principal sensor of DNA DSBs, the MRN complex. Loss of function of PpMRE11 or PpRAD50 strongly and specifically inhibited gene targeting, whilst rates of untargeted transgene integration were relatively unaffected. In contrast, disruption of the PpNBS1 gene retained the wild-type capacity to integrate transforming DNA efficiently at homologous loci. Analysis of the kinetics of DNA-DSB repair in wild-type and mutant plants by single-nucleus agarose gel electrophoresis revealed that bleomycin-induced fragmentation of genomic DNA was repaired at approximately equal rates in each genotype, although both the Ppmre11 and Pprad50 mutants exhibited severely restricted growth and development and enhanced sensitivity to UV-B and bleomycin-induced DNA damage, compared with wild-type and Ppnbs1 plants. This implies that while extensive DNA repair can occur in the absence of a functional MRN complex; this is unsupervised in nature and results in the accumulation of deleterious mutations incompatible with normal growth and developmen

Human factor guidelines for the design of safe in-car traffic information services

The first version of the “Human factor guidelines for the design of safe in-car traffic information services” was compiled in 2014. In 2016 the guidelines were updated by Connecting Mobility/ DITCM, and the present version is a further update of that version. New systems have been introduced into the marked, and the role of apps on smartphones has increased. This report was updated to include recent developments such as gesture control. The guidelines are aimed at in-car traffic information services

MRE11 and RAD50, but not NBS1, are essential for gene targeting in the moss Physcomitrella patens

The moss Physcomitrella patens is unique among plant models for the high frequency with which targeted transgene insertion occurs via homologous recombination. Transgene integration is believed to utilize existing machinery for the detection and repair of DNA double-strand breaks (DSBs). We undertook targeted knockout of the Physcomitrella genes encoding components of the principal sensor of DNA DSBs, the MRN complex. Loss of function of PpMRE11 or PpRAD50 strongly and specifically inhibited gene targeting, whilst rates of untargeted transgene integration were relatively unaffected. In contrast, disruption of the PpNBS1 gene retained the wild-type capacity to integrate transforming DNA efficiently at homologous loci. Analysis of the kinetics of DNA-DSB repair in wild-type and mutant plants by single-nucleus agarose gel electrophoresis revealed that bleomycin-induced fragmentation of genomic DNA was repaired at approximately equal rates in each genotype, although both the Ppmre11 and Pprad50 mutants exhibited severely restricted growth and development and enhanced sensitivity to UV-B and bleomycin-induced DNA damage, compared with wild-type and Ppnbs1 plants. This implies that while extensive DNA repair can occur in the absence of a functional MRN complex; this is unsupervised in nature and results in the accumulation of deleterious mutations incompatible with normal growth and development

Advances in the production of antibodies in plants

Plants are the oldest biotechnological production system and as eukaryots have advantage over bacteria to modify expressed proteins. Laboratory model plants like tobacco along with crop plants with established logistics for harvest and storage like potatoes or rice are used for production of desired proteins. Current trend is to replace soil grown plants with liquid cultures of plant cells grown under controlled conditions. In this respect the moss Physcomitrella patens besides ability to grow in liquid culture enables due to efficient homologous recombination to modify posttranslational glycosylation to correspond human. For heterologous expression in plants, recombinant, single-chain molecules composed of only variable VL and VH regions of the original antibody forming binding pocket are constructed and expressed. Recombinant scFv antibodies are then used as a part of therapeutic or diagnostic drugs in oncology