ITGB5 mutation discovered in a Chinese family with blepharophimosis-ptosis-epicanthus inversus syndrome

Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is a rare autosomal-dominant genetic disorder, and mutations in the forkhead box L2 (FOXL2) gene are one of the major genetic causes. As this study shows, there are many patients with BPES who do not have FOXL2 mutations, as the screening results in all family members were negative. Using whole-exome sequence analysis, we discovered another possible mutational cause of BPES in integrin subunit beta 5 (ITGB5). The ITGB5 mutation (c.608T>C, p.Ile203Thr) appears in the base sequence of all BPES+ patients in this family, and it appears to be a three-generation-inherited mutation. It can cause changes in base sequence and protein function, and there may be cosegregation of disease phenotypes. ITGB5 is located on the long arm of chromosome three (3q21.2) and is close to the known pathogenic gene FOXL2 (3q23). This study is the first to report ITGB5 mutations in BPES, and we speculate that it may be directly involved in the pathogenesis of BPES or indirectly through the regulation of FOXL2.

Potential of Nanoparticles Chitosan for Delivery pcDNA3.1-SB3-HBcAg

Hepatitis B virus (HBV) is a DNA virus that causes hepatitis in humans. This study aims to prepare a Hepatitis DNA vaccine. he optimized base sequence of the SB3-HBcAg gene was derived from the nucleotide base sequence of the Hepatitis B core antigen B3 HBcAg subgenotype, and then Cloning of the pcDNA3.1-SB3-HBcAg has been successfully performed on E. coli DH5α and confirmed by PCR, restriction analysis and sequencing. The propagated plasmids were prepared as DNA-chitosan complex and physiochemical characterized using Particle Size Analyzer. Complex with a 4:1 (wt/wt) ratio of DNA with 0.04% concentration and chitosan have a mean diameter of 231.7 nm and zeta potential +12.3 mV and the value of Cytotoxicity Assay 80-90% as compared to the untreated cells that used as negative control, so it can be concluded that nanoparticles chitosan has good potential as a carrier agent for pcDNA3.1-SB3-HBcAg.

Susceptibility to South African cassava mosaic virus is Associated with a RING Finger E3 Ubiquitin Ligase

BackgroundThe ubiquitylation of proteins is reprogrammed by plant geminiviruses which alter the ubiquitin proteasome system (UPS) to fully infect the host. A RING Finger E3 Ubiquitin Ligase (MeE3L) is located on a major cassava mosaic disease resistance-associated quantitative trait locus. Here, we examine the genetic structure and relative expression of MeE3L (native and gene-edited mutant), and determine how MeE3L affects geminivirus South African cassava mosaic virus (SACMV) DNA accumulation. MethodsCassava protoplasts of model, susceptible and tolerant genotypes were transformed with SACMV infectious clones and/or a CRISPR-editing construct targeting the MeE3L using PEG4000-mediated transfection. DNA and RNA were extracted from transformed protoplasts at 24 hours post-transfection. Relative SACMV load quantitation was determined using DpnI-digested total DNA via qPCR and MeE3L relative expression was determined via reverse transcriptase qPCR, and results were analysed using the 2-ΔΔ method. The MeE3L exonic region was sequenced on the ABI 3500XL Genetic Analyzer platform; and sequences were analysed for mutations and for construction of a phylogenetic tree using the Maximum Likelihood method and Tamura-Nei model.ResultsResults show that SACMV DNA accumulation is cassava genotype-dependent. The study also reveals that native and mutant MeE3L is differentially expressed during SACMV infection in protoplasts of susceptible and tolerant cassava landraces. The susceptible cassava landrace encodes a RINGless MeE3L and the MeE3L base sequence is a determinant of cassava’s response to SACMV. Results further show that SACMV silences the MeE3L RING domain in the susceptible and tolerant landraces; and specifically targets the tolerant MeE3L gene homolog for silencing. ConclusionsThese findings suggest that MeE3L is a target of SACMV, contributing to susceptibility in cassava. The MeE3L base sequence is a determinant of cassava’s response to SACMV. The MeE3L RING domain is actively silenced by SACMV and therefore may be essential for host defence against geminiviruses. The study provides further evidence, in addition to existing literature, that plant E3 ligases are exploited by geminiviruses to enhance pathogenicity.

DNA: Up Close and Personal

We now look at personal genetics and genomics, especially important with the rise of companies willing to analyze your own DNA (for a small fee, of course), giving you the raw genetic information about yourself and your ancestors. Although we previously learned that DNA is “the same” in all species, we now turn to the individual, you, and explore how your DNA base sequence differs from the DNA base sequence of a bacterium, a liverwort, a chimp, and your weird Uncle Jason. This chapter provides the background to appreciate the specific issues related to medical and health issues, and then genealogical studies, coming up in later chapters. For most people, personal genomics testing involves sending a sample of DNA, in the form of spit or a cheek swab, to a lab. What kind of analyses do the labs perform, and what information do they reveal? In addition to full DNA sequence tests, there’s a whole gamut of other DNA tests, including SNP tests, Y-chromosome tests, mtDNA tests, and more. Your DNA base sequence is a gold mine of information unique to you, and it is entirely yours to discover. Whether you are curious about your medical and health genetics, wish to connect with relatives and build a family tree, or are just fascinated at what information your ancestors provided you, these next chapters will help you dig up the hidden secrets of your own genetic heritage.

Genetic Engineering, GMOs, and Genome Editing

DNA is the very core of human existence. The thought of humans manipulating the DNA base sequence of a living thing can be unsettling, disturbing, and sometimes intensely controversial. What are some of the techniques and what are some of the purposes? And what are the concerns? Chapter 10 considers the most controversial use of DNA technology: genetic engineering. It also explores twenty-first century technologies recently developed beyond the “old-fashioned” genetic engineering methods of the 1970s and ’80s. These newer technologies, with curious names, will soon be responsible for putting new products on the market. Synthetic DNA and gene drive are recent additions raising both exciting new possibilities and, simultaneously, old fears. New genome editing technologies, with cool names such as CRISP-Cas9, RNAi, Zinc Finger, and Talens, alter the native DNA in the genome—hence genome editing—and thus forego the need to add DNA from other species or to synthesize entirely. This strategy, say proponents, should quiet the concerns raised from those worried about introducing “foreign” genes from different species. Are you ready?

Your DNA Reveals Medical and Health Surprises

Why are so many people getting their DNA tested? Apart from the science nerds who are always up for such activities, there are two main reasons: health and genealogy. And for each of these there are subgroups. Traditional genealogists hit the proverbial “brick wall” and seek some means to break through, while some adoptees, desperate to find biological family, seem willing to try almost anything. On the other hand, those seeking medical information may have a family history of some frightening health condition, or—due to missing family histories—are in the dark about potential medical issues and want to find out. This chapter first explores personal genomics: the medical and health information held in your DNA base sequence, how to interpret that information, and what may be next on the horizon. What does all this data mean? Can it answer questions such as “Am I carrying around a ticking cancer bomb in my DNA, waiting for me to smoke one more cigarette, or eat one more hot dog before it activates a malignant tumor?”

Sequence-based engineering of dynamic functions of micrometer-sized DNA droplets

DNA has the potential to achieve a controllable macromolecular structure, such as hydrogels or droplets formed through liquid-liquid phase separation (LLPS), as the design of its base sequence can result in programmable interactions. Here, we constructed “DNA droplets” via LLPS of sequence-designed DNA nanostructures and controlled their dynamic functions by designing their sequences. Specifically, we were able to adjust the temperature required for the formation of DNA droplets by designing the sequences. In addition, the fusion, fission, and formation of Janus-shaped droplets were controlled by sequence design and enzymatic reactions. Furthermore, modifications of proteins with sequence-designed DNAs allowed for their capture into specific droplets. Overall, our results provide a platform for designing and controlling macromolecular droplets via the information encoded in component molecules and pave the way for various applications of sequence-designed DNA such as cell mimics, synthetic membraneless organelles, and artificial molecular systems.