what is the effect of asn oligomerization on luvs what is the effect of asn oligomerization on luvs

Written informed consent was obtained from the parents. Once the reaction was stopped by adding 1.33% glycerol and 233mM Tris-HCL (pH8.5), cDNA was purified as above and then subjected to biotinylation by incubating with 83.3mM NaOAc (pH6.0) 0.83mM Biotin hydrazide for 2h at 23C. Genes with expression >=5 CPM in at least two CAGE-Seq libraries (negative control ASOs (A or B)+reference CAGE-Seq libraries) were defined as expressed genes and were tested for DE using EdgeR (ver. Ile and Val (the beta-branched residues) induced the three-stranded oligomerization state, whereas Tyr, Lys, Arg, Orn, Glu and Asp induced the two-stranded state. Mol Cell. Post author: Post published: June 7, Correlation between the effects of ASYN mutations on oligomerization and inclusion formation. Our results showed a thermodynamic stability order (in order of decreasing stability) of: Leu, Met, Ile, Tyr, Phe, Val, Gln, Ala, Trp, Asn, His, Thr, Lys, Ser, Asp, Glu, Arg, Orn, and Gly. Gao F, Cai Y, Kapranov P, Xu D. Reverse-genetics studies of lncRNAs-what we have learnt and paths forward. scared i have esophageal cancer. After purification with the Agencourt RNAClean XP kit (Beckman Coulter), cDNA biotinylation was performed as follows. All Rights Reserved. To characterize this oligomerization and its potential role in GFP transport, cytosolic and secretory forms of enhanced GFP (EGFP) were expressed in GH4C1 and AtT-20 endocrine cells. Ransohoff JD, Wei Y, Khavari PA. By using this website, you agree to our Mol Genet Metab. Orientation, positional, additivity, and oligomerization-state effects of interhelical ion pairs in alpha-helical coiled-coils. Chromatin signature reveals over a thousand highly conserved large non-coding RNAs in mammals. For instance, observed molecular phenotypes must be corroborated by in vitro cellular assays using ASO and orthogonal techniques, such as short interference RNAs (siRNAs) and/or CRISPR interference (CRISPRi). 2015;22(1):57. https://doi.org/10.1038/nsmb.2942. Only samples with a knockdown efficiency higher than 50% in both replicates were subjected to CAGE-Seq. BMC Biol BioMed Central. However, based on the current experimental settings, we cannot completely exclude that some of these changes are also due to impacts intrinsically connected to both negative control ASOs. Weba) By binding to LUVs, the membrane repels oligomers b) By entering the LUVs, it expels their content C) By binding to the oligomers, it blocks membrane permeabilization d) By ! For CAGE-Seq, on the other hand, KD efficiencies were estimated from normalized count per million (CPM) values. oligomer: [noun] a polymer or polymer intermediate containing relatively few structural units. The overall stability range was 7.4 kcal/mol from the lowest to the highest analog, indicating the importance of the hydrophobic core and the dramatic effect a single substitution in the core can have upon the stability of the protein fold. government site. Excitation, emission, and sensitivity were set to 355nm, 460nm, and 14, respectively. Hirakawa S, Hong Y-K, Harvey N, Schacht V, Matsuda K, Libermann T, et al. At passage 7, 7105 LECs and 6105 BECs were seeded into 10cm dishes and cultured overnight. Negative control A or B-specific DE genes were defined as log2FC>1 and FDR<0.05 in negative control A or B and log2FC<1 in negative control B or A for upregulated genes; and log2FC<1 and FDR<0.05 in negative control A or B and log2FC>1 in negative control B or A for downregulated genes (Additional file 4). Molecular modelling studies, however, showed that packing effects could explain deviations from a direct correlation. https://doi.org/10.1186/gb-2013-14-4-r36. An adjective form is oligomeric. Michael Detmar. The https:// ensures that you are connecting to the In particular, we revealed that specifically in LECs, the negative control B exerted off-target effects that included genes associated with LEC biology. Subsequent efforts revealed that the lncRNA family constitutes approximately 72% of the transcribed genome [4]. Codes used to perform differential expression analyses of CAGE-Seq data. An oligonucleotide is a short single-stranded fragment of nucleic acid such as DNA or RNA, or similar fragments of analogs of nucleic acids such as peptide nucleic acid or Morpholinos. Quality control of knockdown efficiencies after lncRNA knockdown. Nonimmune antibody interactions of Group A Streptococcus M and M-like proteins. ASN, bad weather effect on the aircraft. To release 5 cDNA, beads were incubated twice with release buffer (1x RNaseONE buffer (Promega), 0.01% Tween20) for 5min at 95C. Nat Commun Nature Publishing Group. Remember that you can check any relevant video on the PC screen. Abstract. However, all animal aSN oligomers showed weakened permeabilization toward anionic lipid vesicles, indicative of decreased cytotoxicity. Amatschek S, Kriehuber E, Bauer W, Reininger B, Meraner P, Wolpl A, et al. Nature. official website and that any information you provide is encrypted Besides that, we also suggest including alternative negative control ASOs such as mismatched sequences that abrogate the binding to the target sequence. 2021;12(1):9252. 1b, refer to the original study [18] for further details). (a-d) Comparison of knockdown efficiencies after knockdown of 2 LEC and 2 BEC lncRNAs using either negative control A or B, as determined by qPCR (a, b) or CAGE-Seq (c, d). The positive control for the gram stain is? An official website of the United States government. Asn, Gln, Ser, Ala, Gly, Phe, Leu, Met and Trp analogs were indiscriminate and populated two-stranded and three-stranded states. Correspondence to An M protein coiled coil unfurls and exposes its hydrophobic core to capture LL-37. Google Scholar. Despite their common developmental origin, they present distinct functional fates in adulthood that rely on robust lineage-specific regulatory programs. Kornienko AE, Guenzl PM, Barlow DP, Pauler FM. Information and translations of oligomerization in the most comprehensive dictionary definitions resource on the web. 2019;73(5):86983. 2018 Herald International Research Journals. In chemistry and biochemistry, an oligomer (/lmr/ (listen)) is a molecule that consists of a few repeating units which could be derived, actually or conceptually, from smaller molecules, monomers. Moreover, some lncRNA genes do not function through their transcribed RNA molecules but rather through their simple act of transcription [11,12,13,14]. LECs were then transfected with 20nM of negative control A or B and 1L Lipofectamine RNAiMAX previously mixed in 100L Opti-MEM according to the manufacturers instructions. The choice of negative control antisense oligonucleotides dramatically impacts downstream analysis depending on the cellular background. Despite their common If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. Mary Ann Liebert, Inc., publishers 140 Huguenot Street, 3rd Floor New Rochelle, NY 10801 USA. contributed to the in vitro proliferation analysis and provided comments to the manuscript. The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). CAS https://doi.org/10.1038/nrm.2017.104. The oligomerization of such enzymes has been shown to enhance the protein stability and approve allosteric cooperativity to form multiple reaction compartment and provide several interaction site to implement specific function. What is the effect of aSN oligomerization on LUVs? Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. https://doi.org/10.1093/nar/gkm226. 1a). Remember that you can check any relevant video on the PC screen. The liquid oligomerization feed stream is transported upwardly in the reactor vessel against gravity through a fixed bed of solid oligomerization catalyst under really need to submit this soon! 2010;26:13940. 1a). Webwhat is the effect of asn oligomerization on luvs This is a single blog caption. The effect of the extra methylene group on the ligation properties of glutamic (Glu) vs. aspartic (Asp) acid, and glutamine (Gln) vs. asparagine (Asn) amino acids-two pairs of After overnight incubation, cDNA was purified again and subjected to shrimp alkaline phosphatase (1U, Affymetrix) for 30min at 37C. Despite these minor differences, we concluded that both negative control ASOs were suitable for determining the ASO-mediated knockdown efficacy in both cell types. Front genet. The processed data are available at the following link: https://fantom.gsc.riken.jp/6/datafiles/. https://doi.org/10.1038/s41556-019-0311-8. You'll get a detailed solution from a subject matter expert that helps you learn core verificar licencia de conducir venezolana; polish akms underfolder; hhmi biointeractive exploring The AsnAsn interactions work like glue by sticking the DFNKF strands together and assist the on-pathway oligomerization. WebExcessive release of ASN, its oligomerization, aggregation, and deposition in the cytoplasm contribute to neuronal injury and cell death through oxidative-nitrosative stress Increasing evidence suggests that the pathogenicity of -synuclein is correlated with the activity of oligomers formed in the early stages of its aggregation process. Compared to the negative control, what is the effect of aSN monomers on LUVs? The vasculature, composed of blood and lymphatic vessels, undergoes an intricate series of regulatory mechanisms to safeguard the physiological functioning of the organism. The lymphatic and the blood vasculature are closely related systems that collaborate to ensure the organisms physiological function. 2007;109(11):477785. Accessibility In humans, the A53T mutation induces early onset PD and increases the level of aSN oligomerization and fibrillation propensity, but Thr53 occurs naturally in aSNs of most animals. By binding to the oligomers, it blocks membrane permeabilization. Article Cytotoxicity is Asn, Gln, Ser, Ala, Gly, Phe, Leu, Met and Trp analogs were indiscriminate and populated two-stranded and three-stranded states. Biological functions of lymphatic vessels. Myers KJ, Dean NM. Am J Pathol. Given this enrichment on cell proliferation-related terms, we then analyzed empirically whether negative control B was affecting the ability of LECs to proliferate. Micrococcus luteus (Coccus) Clostridium botulinum (Rod) Rhodospirillum rubric (Spiral) What bacteria are used for the gram stain? Nadanaka S, Kagiyama S, Kitagawa H. Roles of EXTL2, a member of the EXT family of tumour suppressors, in liver injury and regeneration processes. M.T., M.I., N.K., and H.S. Third, a rigorous evaluation of dose-response and time-course experiments will help determine the best experimental conditions and provide direct comparisons between experiments [30]. doi: 10.7554/eLife.77989. (e, f) Correlation of knockdown efficiencies between negative control A and B, as determined by qPCR (e) and CAGE-Seq (f). Thus, these results will aid in the de novo design of new coiled-coil structures, a better understanding of their structure/function relationships and the design of algorithms to predict the presence of coiled-coils within native protein sequences. Once the oligomers are formed, you will further purify the sample to separate aSN monomers from oligomers using size exclusion chromatography. (b) Bioinformatic pipeline highlighting the quality control steps prior to the transcriptome profiling after lncRNA candidate knockdowns. Hilton Head Hospital Medical Records, A screening assay for inhibition of Rev binding to a nucleic acid substrate, comprising contacting an isolated nucleic acid comprising two operatively linked binding sites for We used the Model for Oligomerization and Decomposition in Anatomy and Physiology. WebEmployee portals. Before amyloid The oligomeric state(s) was determined by high-performance size-exclusion chromatography and sedimentation equilibrium analysis in benign medium. 5 Ratings, (25 Votes) The growth rate of atmospheric nanoparticles may be affected by particle-phase oligomerization and decomposition of condensing organic molecules. Apr 10 2022 06:11 AM. Will [3] Telomerization is an oligomerization carried out under conditions that result in chain transfer, limiting the size of the oligomers. In f and g, data are displayed as mean values + SD (n=2 in f and n=5 in g). It is thought that one of strategies to prevent and treat PD is inhibition of ASN oligomerization . PLoS Pathog. a) Membrane duplication 2009;38(1):e3. Taken together, these observations support the hypothesis that VA RNAI exerts its inhibitory effect on the NLRP3 inflammasome through binding to PKR. 2013;454(1):13345. First, we recommend including not only libraries from untreated cells but also lipofectamine-only treated cells to evaluate the potential effects of the transfection reagent on the cell of interest [31]. Thus, we pinpoint the essential need to accurately examine multiple negative control ASOs in order to select a proper control set with no to minimal effects on the transcriptome of the targeted cell types. Hon C-C, Ramilowski JA, Harshbarger J, Bertin N, Rackham OJL, Gough J, et al. 2). Nucleic Acids Res. (Select all that apply.) Provided by the Springer Nature SharedIt content-sharing initiative. 2020;21(1):9323. In conclusion, the present study analyzed the effects of negative control ASOs on the transcriptome of LECs and BECs. Geisler S, Coller J. RNA in unexpected places: long non-coding RNA functions in diverse cellular contexts. All these effects induce cell apoptosis . Primers are listed in Additionalfile5. We also advise choosing at least three ASOs targeting the lncRNA candidates that show similar knockdown efficiencies. WebTerms in this set (54) What are the controls for the 3 bacterial shapes? The units of an oligomer may be arranged in a linear chain (as in melam, a dimer of melamine); a closed ring (as in trioxane, a cyclic trimer of formaldehyde); or a more complex structure (as in tellurium tetrabromide, a tetramer of .mw-parser-output .template-chem2-su{display:inline-block;font-size:80%;line-height:1;vertical-align:-0.35em}.mw-parser-output .template-chem2-su>span{display:block;text-align:left}.mw-parser-output sub.template-chem2-sub{font-size:80%;vertical-align:-0.35em}.mw-parser-output sup.template-chem2-sup{font-size:80%;vertical-align:0.65em}TeBr4 with a cube-like core). Interestingly, COLEC12 has been previously reported as a novel lymphatic endothelial cell marker, further supporting the cell type-specific effect of negative control B in LECs [26, 27]. The structure of each analog was analyzed by CD spectroscopy and their relative stability determined by chemical denaturation using GdnHCI (all analogs denatured from the two-stranded state). Report at a scam and speak to a recovery consultant for free. Two types of transformation may occur: true oligomerization and conjunct polymerization. However, very few studies were undertaken to elucidate the role of long noncoding RNAs (lncRNAs) in LEC and BEC biology. Gapmers: methods and protocols. 3e), primarily related to chromatin organization and endothelial cell proliferation. COLEC12 regulates apoptosis of osteosarcoma through Toll-like receptor 4activated inflammation. [5] Different heterogeneous and homogeneous catalysts are operative in producing green oils via the oligomerization of alkenes.[6]. For instance, previous studies showed the possibility of delivering ASOs by gymnosis [38, 39]. RNA maps reveal new RNA classes and a possible function for pervasive transcription. C.-C.H. RNase I treatment was performed on purified cDNA samples using 5units (U) RNase ONE ribonuclease (Promega) for 30min at 37C. Manage cookies/Do not sell my data we use in the preference centre. At each time point, LECs were washed with DPBS (Thermo Fisher Scientific), and 100L of 0.1mg/mL MUH (Sigma) in DPBS were added to each well. This finding was further supported by correlation analysis between negative control A and B KD efficiencies, where a lower but still significant correlation was observed in CAGE-Seq data in comparison to qPCR results (Fig. 2d). which part of the P-values were calculated using linear regression. Nat Rev Genet Nature Publishing Group. (g) Quantification of the 4-methylumbelliferyl heptanoate (MUH) proliferation assay over 72h in neonatal LECs derived from the same donor after negative control A or B transfection. In general, the side-chain contribution to the level of stability correlated with side-chain hydrophobicity. Specific expression patterns of lncRNA subsets have also been associated with cell state coordination, cell differentiation, development, and disease progression [15, 16]. Azobenzene-Based Amino Acids for the Photocontrol of Coiled-Coil Peptides. 1a). 22 Jun. herald express torbay family announcements which part of the chromatogram represents the asn oligomers The Asn-Asn stacking observed here is similar to the Asn ladder commonly found in globular -helix proteins. Disclaimer. Cite this article. a) Last updated: 7/23/2022 What is the effect of aSN oligomerization on LUVs? In regards to oligomerization state, eight analogs demonstrated the ability to populate exclusively one oligomerization state in benign buffer (0.1 M KCl, 0.05 M K(2)PO(4)(pH 7)). The Nutrizion > Blog Classic > Uncategorized > what is the effect of asn oligomerization on luvs. Anatomy and Physiology questions and answers. Ligated cDNA was purified again and subjected to second-strand synthesis by incubating with 1x ThermoPol reaction buffer pack (New England Biolabs), 0.2mM dNTPs, 1mM nAnT-iCAGE 2nd primer, 2U DeepVent (exo-) DNA pol (New England Biolabs) for 5min at 95C, 5min at 55C, and 30min at 72C. L.D. However, we also observed that referencing to either negative control A or B led to slight differences in the degree of the KD efficiencies in our CAGE-Seq data compared to qPCR results (Fig. Groundhog Max Problems, container_memory_working_set_bytes vs container_memory_usage_bytes, National Financial Services Llc Albuquerque, Nm, johnson funeral home anderson sc obituaries. WebAntisense oligonucleotide (ASOs) are small-sized single-stranded nucleic acids and offer some advantage over siRNAs in terms of targeting both nuclear and cytoplasmic located Mercer TR, Dinger ME, Mattick JS. https://doi.org/10.1093/nar/gky437. 2.0.12) [43] applying default settings (Additional file 2). https://doi.org/10.1016/j.ymgme.2017.10.004. The model protein consists of two identical 38 residue polypeptide chains covalently linked at their N termini via a disulfide bridge. Trends Genet. WebOligomerization of protein complexes has been involved in various mechanisms that play a major role in endoplasmic reticulum (ER) functions. This mobility is a function of the length, conformation, and charge of the molecule. doi: 10.1371/journal.ppat.1009248. Oncotarget Impact J. Purified total RNA (4g) was first subjected to reverse transcription using anchored random primers and Superscript III reverse transcriptase (Thermo Fisher Scientific) for 30s at 25C and 1h at 50C. Cytotoxicity is mainly associated with the oligomeric species, but we still know little about their assembly and structure. While Mic10 is part of a large hetero-oligomeric complex, it has been proposed that the protein itself might form homo-oligomers in the IM (Harner et al., 2011).Mic10 has two highly conserved elongated GxxxG motifs in its two transmembrane domains (Figure After removing low-quality sequencing reads, each library contains, on average, a total of 15 million reads. Robinson MD, McCarthy DJ, Smyth GK. 3g). The recent technological boost in sequencing approaches unveiled long noncoding RNAs (lncRNAs) as prominent regulatory players of various gene expression levels in a cell-type-specific manner. MeSH The term multimer (/mltmr/) is used in biochemistry for oligomers of proteins that are not covalently bound. a) None, they remain the same b) Conformational control in a photoswitchable coiled coil. Increasing evidence suggests that the pathogenicity of -synuclein is correlated with the activity of oligomers formed in the early stages of its aggregation process. Yao RW, Wang Y, Chen L-L. Cellular functions of long noncoding RNAs. Methods Mol Biol Springer, New York, NY. tRNA-coated magnetic beads were then washed twice with wash buffer A (4.5M NaCl, 50mM EDTA (pH8.0), 0.1% Tween20), and resuspended in wash buffer A containing 3.75g tRNA. Protein oligomerization may be an advantageous feature from the perspective of protein evolution for a number of reasons, including new opportunities for functional control, such as allosteric regulation and the establishment of higher-order complexity. Robinson MD, Oshlack A. A key feature of Parkinson disease is the aggregation of -synuclein and its intracellular deposition in fibrillar form. 2a, b), saturation concentrations of oligomerization product (C p *; Fig. Download scientific diagram | Reduction of Ab 42 oligomerization: (a and b) the effect of SP1 on LUV dye leakage caused by Ab 42 aggregates (incubated at varied time intervals: 1 Features. After alignment, the expression for CAGE-Seq promoters was estimated as previously described [20]. In addition to that, the increasing evidence that lncRNAs are involved in various aspects of gene expression regulation emphasizes the relevance of lncRNA classification based on their functions [9, 10]. 1999 Jan 15;285(2):785-803. doi: 10.1006/jmbi.1998.2284. HHS Vulnerability Disclosure, Help J Mol Biol. Throughout this work, we report results for the oligomerization of ethylene over Ni-H in a packed bed reactor. Then, 2U USER enzyme (New England Biolabs) were added to the SAP-treated cDNA and further incubated for 30min at 37C followed by 5min at 95C. 2014;1164:6785. 2021 Feb 25;17(2):e1009248. Vasquez G, Freestone GC, Wan WB, Low A, De Hoyos CL, Yu J, et al. The results showed that perturbation using negative control A and B caused the deregulation of 744 (up: 430; down: 314) and 813 (up: 454; down: 359) genes in LECs and 2487 (up: 1371; down: 1116) and 2487 (up: 1383; down: 1104) in BECs (Additionalfile3). 2011;12(12):86174. Part of 3a, b and Additionalfile4), which is likely to be attributable to the lipofectamine treatment as previously observed in human dermal fibroblasts [20]. Here, we describe the quality control steps adopted in our analysis pipeline before determining the knockdown effects of three ASOs per lncRNA target on the LEC or BEC transcriptomes. Available from: http://www.bioinformatics.babraham.ac.uk/projects/fastqc/. https://doi.org/10.1038/s41587-019-0106-2. The FC values of the DE genes were largely overlapping between both negative control ASOs (Fig. After an additional purification step, cDNA concentration was measured using Quant-iT OliGreen ssDNA reagent and kit (Thermo Fisher Scientific), and the ratio of mRNA/rRNA was analyzed by performing qPCR with ACRB-specific primers and 18S ribosomal cDNA primers on a 7900HT real-time system (Applied Biosystems). 2019;37(6):64050. Genetic analysis of protein stability and function. https://doi.org/10.1038/s41467-021-21217-0. 2010;11(3):R259. FOIA Therapeutic silencing of an endogenous gene by systemic administration of modified siRNAs. Protein oligomerization may often be an advantageous feature from the perspective of protein evolution and has probably evolved by a variety of mechanisms. This alteration eventually lead to widespread neuronal death and dementia [23] . DNA digestion was performed using the RNase-free DNase set (Qiagen). 2009;10(3):1559. Oligomers toxicity seems to be associated with both their ability to bind and affect the Cytotoxicity is attributed to a soluble aSN oligomeric species that permeabilizes membranes significantly more than monomers and fibrils. The lymphatic and the blood vasculature are closely related systems that collaborate to ensure the organisms physiological function. What is the effect of aSN oligomerization on LUVs? which part of the chromatogram represents the asn oligomers which part of the chromatogram represents the asn oligomers The effects of different parameters on the growth with oligomerization can be seen in Fig. For other uses, see, Srpskohrvatski / , "Glossary of basic terms in polymer science (IUPAC Recommendations 1996)", "Heterogeneous catalysts for gas-phase conversion of ethylene to higher olefins", https://en.wikipedia.org/w/index.php?title=Oligomer&oldid=1140991496, Short description is different from Wikidata, Creative Commons Attribution-ShareAlike License 3.0, This page was last edited on 22 February 2023, at 20:21. This site needs JavaScript to work properly. In this regard, we especially observed that the choice of negative control ASOs can dramatically impact the conclusions drawn from the analysis depending on the cellular background. SCORE: 82 / 120 DAY 2; 23:23 PROGA Click to answer the quiz a HOME THEORY MEDIA MISSION What is the effect of aSN oligomerizatio. Genome Res Cold Spring Harbor Lab. National Library of Medicine 1b). which part of the chromatogram represents the asn oligomersst charles county courthouse auctionsst charles county courthouse auctions In the process, a liquid oligomerization feed stream comprising lighter aliphatic olefins is passed to a reactor vessel. What is the effect of aSN oligomerization on LUVs? WebInducing and purifying aSN oligomers After purifying the aSN monomers, you will induce the formation of aSN oligomers using a simple heating procedure. (a, b) Correlation of log2FC between differentially expressed (DE) genes in negative control A and B. Separation of Coiled-Coil Structures in Lamin A/C Is Required for the Elongation of the Filament. In our recent studies, we efficiently connected the molecular phenotypes associated with the lncRNA target knockdown, as predicted by the CAGE-Seq data analysis, with essential cellular functions and provided detailed evidence on their molecular mode of actions combining RNA-DNA, RNA-protein, and RNA-chromatin interaction studies [18, 20]. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Federal government websites often end in .gov or .mil. Would you like email updates of new search results? Single-stranded cDNA was then purified with Agencourt AMPure XP kit (Beckman Coulter) and subjected to another RNase I (5U) treatment for 30min at 37C. Second, analysis of gap ablated ASO and ASO backbone modification data can provide further useful information on the off-target effects of the negative control ASOs and to differentiate between off-target cleavage and steric hindrance [34,35,36]. Along with the necessary repetition of the experimental procedure, these guidelines will help design coherent lncRNA knockdown studies leading to a solid interpretation of lncRNA knockdown effects. Bison can run at speeds of up to 70 After exonuclease I (20U, New England Biolabs) digestion for 30min at 37C, purified cDNA sample quality was assessed for linker dimers using Agilent Bioanalyzer (Agilent Technologies), and its concentration was measured using Quant-iT PicoGreen dsDNA reagent and kit (Thermo Fisher Scientific). 1998 Nov 13;283(5):993-1012. doi: 10.1006/jmbi.1998.2125. Hotait M, Nasreddine W, El-Khoury R, Dirani M, Nawfal O, Beydoun A. FARS2 mutations: more than two phenotypes?