• Title/Summary/Keyword: Base substitution

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Transition Substitution of Desired Bases in Human Pluripotent Stem Cells with Base Editors: A Step-by-Step Guide

  • Ju-Chan Park;Keun-Tae Kim;Hyeon-Ki Jang;Hyuk-Jin Cha
    • International Journal of Stem Cells
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    • v.16 no.2
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    • pp.234-243
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    • 2023
  • The recent advances in human pluripotent stem cells (hPSCs) enable to precisely edit the desired bases in hPSCs to be used for the establishment of isogenic disease models and autologous ex vivo cell therapy. The knock-in approach based on the homologous directed repair with Cas9 endonuclease, causing DNA double-strand breaks (DSBs), produces not only insertion and deletion (indel) mutations but also deleterious large deletions. On the contrary, due to the lack of Cas9 endonuclease activity, base editors (BEs) such as adenine base editor (ABE) and cytosine base editor (CBE) allow precise base substitution by conjugated deaminase activity, free from DSB formation. Despite the limitation of BEs in transition substitution, precise base editing by BEs with no massive off-targets is suggested to be a prospective alternative in hPSCs for clinical applications. Considering the unique cellular characteristics of hPSCs, a few points should be considered. Herein, we describe an updated and optimized protocol for base editing in hPSCs. We also describe an improved methodology for CBE-based C to T substitutions, which are generally lower than A to G substitutions in hPSCs.

Mutational Analysis of the Region between vir Box and -35 Sequence in virE Promoter of pTiA6

  • Woong Seop Sim
    • Journal of Plant Biology
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    • v.38 no.3
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    • pp.259-266
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    • 1995
  • To investigate how the dyad symmetry region (DSR) and the distance between vir box and -35 sequence of the virE promoter plays a role in virE gene expression, two mutants were constructed by base substitution and insertional mutagenesis. The base substitutional mutation, a AAlongrightarrowCG substitution at positions -39 and -40 on the DSR, showed the level of $\beta$-galactosidase activity approximately 91% of the wild type virE promoter activity. Therefore, the native structure of the DSR seems to be not essential for virE expression. The insertional mutation, constructed by inserting 8 bp ClaI linker between -49 and -50, displayed the $\beta$-galactosidase activity at 12% of the native virE promoter activity. However, this striking reduction appears to be not caused by destruction of the native DSR structure, but by shifting the vir box far from putative -35 sequence.

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Mutagenicity of Chinese Herbal Anti-cancer Drugs and Their Antimutagenic Activity to Base-pair Substitution Mutagen (치암중초약의 돌연변이유발 및 돌연변이유발 억제효과)

  • Lee, Hyun-Chul
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.2
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    • pp.185-193
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    • 1987
  • Ten species of herbae, which have been used to treat cancers in Chinese medicine, were tested to investigate their mutagenicity or antimutagenicity in S. typhimurium TA97, TA98, TA100, TA1535, and TA1538. Scolopendra centipede was weakly active in reversion of the frameshift mutation in S. typhimurium TA97 strain and the base-pair substitution in TA100 and TA1535 strains. Other herbae such as Coix lachryma, Dianthus superbus, Tricanthoshse kirilowii, Eupatorium formosanum, Lithospermum erythrorhizon, Ansaema japonicum, Curcuma zedoaria, Helicteres angustifolia, and Euonymus sieboldianus did not show any of the mutagenic potential, regardless of the metabolic activation with rat hepatic microsomal fraction. Dianthus superbus, Eupatorium formosanum, and Euonymus sieboldianus exhibited suppressive activities on microbial mutagenesis of N-methyl-N'-nitrosoguanidine, a base-pair substitution mutagen, in TA1535 and TA100 tester strains. The antimutagenic activities of Dianthus superbus and Euonymus sieboldianus appeared to be dose-dependant.

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Study on Quality of Yukwa by Substitution with Resistant Starch (저항전분 대체에 따른 유과의 품질에 관한 연구)

  • Lee, Mi Hye;Oh, Myung Suk
    • Journal of the East Asian Society of Dietary Life
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    • v.26 no.5
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    • pp.407-417
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    • 2016
  • Effects of resistant starch as a functional substitute on the quality of Yukwa were determined. Replacement ratios of resistant starch were 2, 4, and 6% of glutinous rice flour. Regarding pasting properties of Yukwa dough replaced with resistant starch, initial pasting temperature significantly increased and peak viscosity significantly decreased with increasing resistant starch. The moisture contents of bandegi and Yukwa base replaced with resistant starch were higher than that of the control. The expansion ratio of Yukwa base replaced with resistant starch significantly decreased with increasing resistant starch, and there were no significant differences in the oil absorption ratio. Lightness (L) of Yukwa base replaced with resistant starch increased significantly with increasing resistant starch. Appearance of Yukwa base replaced with resistant starch showed a shortened length and increased width with increasing resistant starch, and cross-section showed an oval shape, fine air cell distribution, and increased outer layer thickness. Hardness and peak number increased significantly with increasing resistant starch. The overall acceptability of Yukwa base replaced with 6% resistant starch was the highest among the samples, but the result was not significant. The above results show that resistant starch substitution in Yukwa improved the texture and further could improve health functionality due to its dietary fiber content. Resistant starch was appropriate as a 6% replacement for glutinous rice flour in Yukwa.

An Improved Steganography Method Based on Least-Significant-Bit Substitution and Pixel-Value Differencing

  • Liu, Hsing-Han;Su, Pin-Chang;Hsu, Meng-Hua
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.14 no.11
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    • pp.4537-4556
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    • 2020
  • This research was based on the study conducted by Khodaei et al. (2012), namely, the least-significant-bit (LSB) substitution combined with the pixel-value differencing (PVD) steganography, and presented an improved irreversible image steganography method. Such a method was developed through integrating the improved LSB substitution with the modulus function-based PVD steganography to increase steganographic capacity of the original technique while maintaining the quality of images. It partitions the cover image into non-overlapped blocks, each of which consists of 3 consecutive pixels. The 2nd pixel represents the base, in which secret data are embedded by using the 3-bit LSB substitution. Each of the other 2 pixels is paired with the base respectively for embedding secret data by using an improved modulus PVD method. The experiment results showed that the method can greatly increase steganographic capacity in comparison with other PVD-based techniques (by a maximum amount of 135%), on the premise that the quality of images is maintained. Last but not least, 2 security analyses, the pixel difference histogram (PDH) and the content-selective residual (CSR) steganalysis were performed. The results indicated that the method is capable of preventing the detection of the 2 common techniques.

Sequence Analysis of the Internal Transcribed Spacer of Ribosomal DNA in the Genus Rhizopus

  • Park, You-Jung;Min, Byung-Re
    • Mycobiology
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    • v.33 no.2
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    • pp.109-112
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    • 2005
  • The internal transcribed spacer (ITS) regions including the 3'-end of 18S rRNA gene, 5.8S rRNA gene and the 5'-end of the 28S rRNA gene of Rhizopus spp. were amplified by PCR and analyzed by DNASIS program. Length polymorphism of these region ranged from 564 bp in R. oryzae to 789bp in R. stolonifer. The length and sequence of 5.8S was very conserved with $154{\sim}155\;bp$. The sequence of ITS2 was more variable than that of ITS1. The base substitution rates were ranged from 0 to 0.6069 per site, and higher rate was found in R. stolonifer. In general, transition was usually more frequent than transversion. On the basis of sequencing results, four groups were clustered with value of 61.9% similarity; R. oryzae, R. micros pores, R. homothallicus, and R. stolonifer groups.

Effect of Processing on the Antimutagenicity of Rice (쌀의 돌연변이 억제활성에 미치는 가공처리의 영향)

  • Kim, In-Ho;Chun, Hyang-Sook;Ha, Tae-Youl;Moon, Tae-Wha
    • Korean Journal of Food Science and Technology
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    • v.27 no.6
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    • pp.944-949
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    • 1995
  • To investigate the effect of processing on the antimutagenicity of rice, antimutagenic effect of rice products including cooked rice, plain steamed rice bread(baikseolgi) and parched rice powder were observed. They showed inhibitory effects of $46%{\sim}100%$ on the frameshift mutation, whereas they showed little inhibitory effect on the base substitution mutation in the Salmonella typhimurium reversion assay. No inhibitory effect was found in the SOS chromotest. Inhibitory effect was more clearly observed on the indirect-acting mutagen than on the direct-acting mutagen. In case of the S. typhimurium reversion assay, the range of inhibition rate against the frameshift and base substitution mutation was $75%{\sim}100%$ and $66%{\sim}87%$, respectively, and was $19%{\sim}67%$ in the SOS chromotest. Antimutagenic activity of raw rice was supposed to be maintained in processed rice products.

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Genetic Variation in Exon 3 of Human Apo B mRNA Editing Protein (apobec-1) Gene

  • Hong, Seung-Ho;Song, Jung-Han;Kim, Jin-Q
    • Journal of Genetic Medicine
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    • v.3 no.1
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    • pp.15-19
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    • 1999
  • We have investigated the genetic variation in the human apo B mRNA editing protein (apobec-1) gene. Exon 3 of the apobec-1 gene was amplified by polymerase chain reaction. After detection of an additional band by single strand conformational polymorphism (SSCP) analysis, sequencing of the SSCP-shift sample revealed a single-base mutation. The mutation was a CGG transversion at codon 80 resulting in a lleRMet substitution. This substitution was confirmed by restriction fragment length polymorphism analysis since a Pvull site is abolished by the substitution. Population and family studies confirmed that the inheritance of the genotypes for apobec-1 gene polymorphism is controlled by two codominant alleles (P1 and P2). A significant difference in plasma triglyceride was detected among the different apobec-1 genotypes in the CAD patients (P<0.05). Our study could provide the basis for elucidating the interaction between genetic variation of the apobec-1 gene and disorders related to lipid metabolism.

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Nanocrystallization Behavior of Al-Y-Ni with Cu Additions (Cu 첨가에 따른 Al-Y-Ni의 나노결정화 거동)

  • 홍순직;천병선;강세선;이임렬
    • Journal of Powder Materials
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    • v.9 no.1
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    • pp.19-24
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    • 2002
  • This paper reports the results of an investigation into the effect of Cu additions upon the nano-crystallization behaviour of an Al-Y-Ni alloy. 1 at.% Cu was added to a base alloy of Al/sub 88/Y₄Ni/sub 8/ either by substitution for Al to form Al/sub 87/Y₄Ni/sub 8/Cu₁, or by substitution for Ni to form Al/sub 88/Y₄Ni/sub 7/Cu₁. Consistent with previous findings in the literature, the substitution of Cu for Al was found to increase the thermal stability of the amorphous phase whereas the substitution of Cu for Ni was found to decrease its thermal stability. Comparing the microstructures of these alloys after heat treatment to produce equivalent volume fractions of Al nanocrystals showed average grain sizes of 14 nm, 12 nm and 9 nm for the alloys Al/sub 88/Y₄Ni/sub 8/, Al/sub 87/Y₄Ni/sub 8/Cu₁respectively. The effect of Cu in refining the size of the nanocrystals was attributed to enhanced nucleation increasing the number density of the nanocrystals, rather than diffusion limited or interface limited growth.