当前位置： SCI文献检索 > MOLECULAR BIOLOGY REPORTS期刊下所有文献 > Evaluation of multiple gene targeting in porcine embryos by the CRISPR/Cas9 system using electroporation.

Evaluation of multiple gene targeting in porcine embryos by the CRISPR/Cas9 system using electroporation.

Abstract：

:The CRISPR/Cas9 system now allows for unprecedented possibilities of genome editing. However, there are some limitations, including achieving efficient one-step multiple genome targeting to save costs, time, and ensure high quality. In the present study, we investigated the efficiency of one-step multiple gene modification by electroporation in porcine zygotes using pooled guide RNAs (gRNAs) targeting CMAH, GHR, GGTA1, and PDX1. We first selected the best-performing gRNA from three different designs for each gene based on the effect on embryo development and mutation efficiency. The three gRNAs showed equivalent effects on the rates of blastocyst formation in each targeted gene; however, gRNAs CMAH #2, GHR #3, GGTA1 #3, and PDX1 #3 showed the highest biallelic mutation rate, although the total mutation rate of PDX1 #3 was significantly lower than that of PDX1 #1. Therefore, CMAH #2, GHR #3, GGTA1 #3, and PDX1 #1 were used as a mixture in electroporation to further clarify whether multiple genes can be targeted simultaneously. Individual sequencing of 43 blastocysts at the target sites of each gene showed mutations in one and two target genes in twenty-four (55.8%) and nine (20.9%) blastocysts, respectively. No mutation was detected in any target gene in ten (23.3%) blastocysts and no blastocysts had a mutation in three or more target genes. These results indicate that electroporation could effectively deliver multiple gRNAs and Cas9 protein into porcine zygotes to target multiple genes in a one-step process. However, the technique requires further development to increase the success rate of multiple gene modification.

journal_name

Mol Biol Rep

journal_title

Molecular biology reports

authors

Hirata M,Wittayarat M,Namula Z,Le QA,Lin Q,Nguyen NT,Takebayashi K,Sato Y,Tanihara F,Otoi T

doi

10.1007/s11033-020-05576-3

subject

Has Abstract

pub_date

2020-07-01 00:00:00

pages

5073-5079

issue

7

eissn

0301-4851

issn

1573-4978

pii

10.1007/s11033-020-05576-3

journal_volume

47

pub_type

杂志文章

相关文献

MOLECULAR BIOLOGY REPORTS文献大全
  • Molecular cloning, characterization and differential expression of Cdc42 in Fonsecaea monophora.

    abstract：:The cell divisions cycle 42 (Cdc42) gene has been characterized in the fungi, such as Candida albicans, Penicillium marneffei, and Wangiella (Exophiala) dermatitidis, which plays important roles during growth and development. The partial cDNA sequence of Cdc42 of Fonsecaea monophora (F. monophora), designated FmCdc42,...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-011-0806-2

    authors： Xie Z,Feng P,Zhang J,Li X,Sun J,Lu C,Huang H,Xi L

    更新日期：2012-02-01 00:00:00

  • Expression of NAT2 in immune system cells and the relation of NAT2 gene polymorphisms in the anti-tuberculosis therapy in Mexican mestizo population.

    abstract：:Arylamine N-acetyltransferase 2 (NAT2) metabolizes isoniazid (INH) and Single Nucleotide Polymorphisms (SNP) responsible for its activity has been reported. The aim of this study in the Mexican mestizo population was to evaluate NAT2 expression at the protein level in immune cells, as well as the distribution and freq...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-014-3677-5

    authors： Salazar-González R,Gómez R,Romano-Moreno S,Medellín-Garibay S,Núñez-Ruíz A,Magaña-Aquino M,Milán-Segovia RC,Portales-Pérez DP

    更新日期：2014-12-01 00:00:00

  • RA induces the neural-like cells generated from epigenetic modified NIH/3T3 cells.

    abstract：:Recently, differentiated somatic cells had been reprogrammed to pluripotential state in vitro, and various tissue cells had been elicited from those cells. Epigenetic modifications allow differentiated cells to perpetuate the molecular memory needed for the cells to retain their identity. DNA methylation and histone d...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-009-9489-3

    authors： Zhang XM,Li QM,Su DJ,Wang N,Shan ZY,Jin LH,Lei L

    更新日期：2010-03-01 00:00:00

  • Bsu2413I and Bfi2411I, two new thermophilic type II restriction endonucleases from Bacillus subtilis and Bacillus firmus: isolation and partial purification. Thermophilic endonucleases from two Bacillus species.

    abstract：:Two new thermophilic type II restriction endonucleases, which we designated as Bsu2413I and Bfi2411I, have been isolated from gram-positive thermophilic bacteria Bacillus subtilis strain 2413 and Bacillus firmus strain 2411 respectively and partially purified. The restriction endonucleases were extracted from cell ext...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1023/b:mole.0000031503.19851.3b

    authors： Jutur PP,Hoti SL,Reddy AR

    更新日期：2004-06-01 00:00:00

  • Human dipeptidyl peptidase III mRNA variant I and II are expressed concurrently in multiple tumor derived cell lines and translated at comparable efficiency in vitro.

    abstract：:Dipeptidyl peptidase III (DPP III) is an emerging biomarker of human cancers. Expression, specificity, and function of human DPP III (hDPP III) mRNA variant I (V-I), II (V-II), and III (V-III) are poorly understood. Here, we investigated expression of these variants in multiple human tumor derived cell lines. DNA sequ...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-016-3996-9

    authors： Prajapati SC,Chauhan SS

    更新日期：2016-06-01 00:00:00

  • Oxidative stress induced by high-glucose diet in liver of C57BL/6J mice and its underlying mechanism.

    abstract：:High glycemic index diet can induce multiple diseases. Many research indicated that oxidative stress played important role in many pathological conditions. However, the impact of gene expression and dietary habit on oxidation process are still less clear. We used high-glucose diet to feed C57BL/6J mice for 4 weeks, me...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-010-0039-9

    authors： Du D,Shi YH,Le GW

    更新日期：2010-12-01 00:00:00

  • DNA supercoiling by gyrase is linked to nucleoid compaction.

    abstract：:The genes of E. coli are located on a circular chromosome of 4.6 million basepairs. This 1.6 mm long molecule is compressed into a nucleoid to fit inside the 1-2 microm cell in a functional format. To examine the role of DNA supercoiling as nucleoid compaction force we modulated the activity of DNA gyrase by electroni...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1023/a:1020318705894

    authors： Stuger R,Woldringh CL,van der Weijden CC,Vischer NO,Bakker BM,van Spanning RJ,Snoep JL,Westerhoff HV

    更新日期：2002-01-01 00:00:00

  • Human intracerebroventricular (ICV) injection of autologous, non-engineered, adipose-derived stromal vascular fraction (ADSVF) for neurodegenerative disorders: results of a 3-year phase 1 study of 113 injections in 31 patients.

    abstract：:We have chosen to test the safety of human intracerebroventricular (ICV) brain injections of autologous non-genetically-modified adipose-derived stromal vascular fraction (ADSVF). In this IRB-approved trial, 24 patients received ICV ADSVF via an implanted reservoir between 5/22/14 and 5/22/17. Seven others were inject...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-019-04983-5

    authors： Duma C,Kopyov O,Kopyov A,Berman M,Lander E,Elam M,Arata M,Weiland D,Cannell R,Caraway C,Berman S,Scord K,Stemler L,Chung K,Khoudari S,McRory R,Duma C,Farmer S,Bravo A,Yassa C,Sanathara A,Singh E,Rapaport B

    更新日期：2019-10-01 00:00:00

  • Partial purification and characterization of a double-stranded RNA-specific nuclease from human placenta.

    abstract：:A double-stranded RNA-specific nuclease (ds RNase) has been isolated and partially purified from human placenta by DEAE-cellulose and DNA-cellulose column chromatography. Denatured DNA-cellulose retained most of the single-stranded RNA-specific nuclease (ss RNase) activity, whereas the ds RNase came out in the void vo...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/BF00775365

    authors： Kalyanaraman S,Maran A,Shanmugam G

    更新日期：1983-08-01 00:00:00

  • Molecular insight into the metabolic activities of a protein-rich micro alga, Arthrospira platensis by de novo transcriptome analysis.

    abstract：:To gain genetic insights into the protein-rich microalga, the transcriptome of Arthrospira platensis was sequenced using Illumina technology and de novo assembly was carried out. A total of 6023 transcripts were present in the transcriptome among which 4616 transcripts were annotated with specific functions. Gene onto...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-018-4229-1

    authors： Kumaresan V,Sannasimuthu A,Arasu MV,Al-Dhabi NA,Arockiaraj J

    更新日期：2018-10-01 00:00:00

  • Molecular cloning and characterization of the porcine FTO promoter and coding regions.

    abstract：:The fat mass and obesity associated gene (FTO) has been widely reported to be associated with fat mass or fat deposition in different species. In the present study, we cloned both promoter and coding regions of the gene in pigs with over 5 Kb of sequence for the former region and 1,596 bp for the latter region. Compar...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-010-0431-5

    authors： Huang J,Yang Y,Liu G,Zhang J,Dong X,Bai Y,Fang M

    更新日期：2011-04-01 00:00:00

  • Machine learning algorithm-based risk prediction model of coronary artery disease.

    abstract：:In view of high mortality associated with coronary artery disease (CAD), development of an early predicting tool will be beneficial in reducing the burden of the disease. The database comprising demographic, conventional, folate/xenobiotic genetic risk factors of 648 subjects (364 cases of CAD and 284 healthy controls...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-018-4236-2

    authors： Naushad SM,Hussain T,Indumathi B,Samreen K,Alrokayan SA,Kutala VK

    更新日期：2018-10-01 00:00:00

  • Effect of mechanical stretch on the expressions of elastin, LOX and Fibulin-5 in rat BMSCs with ligament fibroblasts co-culture.

    abstract：:The occurrence of PFD is closely related with elasticity, toughness, and functional changes of the connective tissue of the pelvic support tissue. This study aims to evaluate the effect of mechanical stretch on the differentiation of BMSCs with a co-culture with pelvic ligament fibroblasts. BMSCs was isolated and iden...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-011-1422-x

    authors： Bing Z,Linlin L,Jianguo Y,Shenshen R,Ruifang R,Xi Z

    更新日期：2012-05-01 00:00:00

  • Isolation and characterization of a cotton cdh-like gene.

    abstract：:Cotton fiber cells elongate without dividing to form economically valuable spinnable fiber. Reports of the ploidy level of fiber cells are variable. Early reports indicated an increase in nuclear DNA content in young fibers; however, subsequent reports failed to observe such a significant increase in ploidy level. Eva...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-009-9496-4

    authors： Taliercio E,Ray J,Scheffler J

    更新日期：2010-02-01 00:00:00

  • ZmMYB31, a R2R3-MYB transcription factor in maize, positively regulates the expression of CBF genes and enhances resistance to chilling and oxidative stress.

    abstract：:Maize (Zea mays L.) is an important model plant with an important role in agriculture and national economies all over the world. The optimum growth temperature of maize is between 25 and 28 °C. At temperatures below 12 °C, maize is vulnerable to damage by chilling stress. MYB transcription factors play important roles...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-019-04840-5

    authors： Li M,Lin L,Zhang Y,Sui N

    更新日期：2019-08-01 00:00:00

  • Regulation analysis of contractile ATPase flux in skeletal muscle.

    abstract：:J The [Ca2+] regulation of contractile ATPase flux, Jp, in skeletal muscle was analysed by computation of the Response R(Jp) Ca2+ for a 10 Hz range of electrical stimulation frequencies. Results of our analysis of the kinetic controls in ATP free energy metabolism in a network model of contracting muscle (J.A.L. Jenes...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1023/a:1020317610863

    authors： Jeneson JA

    更新日期：2002-01-01 00:00:00

  • EGY2, a chloroplast membrane metalloprotease, plays a role in hypocotyl elongation in Arabidopsis.

    abstract：:Intramembrane proteases control many important processes in a wide variety of organisms through regulated intramembrane proteolysis (RIP). However, very few intramembrane proteases have been characterized in plants. Intriguingly, EGY2 in Arabidopsis belongs to the Site-2 protease (S2P) family that performs RIP. It con...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-011-0962-4

    authors： Chen G,Law K,Ho P,Zhang X,Li N

    更新日期：2012-03-01 00:00:00

  • Isolation of microsatellite loci for the endangered vermetid gastropod Dendropoma lebeche using Illumina MiSeq next generation sequencing technology.

    abstract：:Dendropoma petraeum, considered the primary vermetid reef-building species in the Mediterranean, has recently been shown to be a species complex of at least four cryptic species. These species have highly restricted, non-overlapping distributions, causing concern for their conservation status. To better study the gene...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-018-4346-x

    authors： López-Márquez V,García-Jiménez R,Calvo M,Templado J,Machordom A

    更新日期：2018-12-01 00:00:00

  • Association between activity and genotypes of paraoxonase1 L55M (rs854560) increases the disease activity of rheumatoid arthritis through oxidative stress.

    abstract：:Rheumatoid arthritis (RA) is considered as a long-term autoimmune disorder. Gene polymorphism and oxidative stress might be involved in the pathogenesis of the disease. We aimed to determine the association between PON-1L55M polymorphism and its effects on inflammatory markers such as anti-cytroline circulated-peptide...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-018-4530-z

    authors： Tanhapour M,Shahmohamadnejad S,Vaisi-Raygani A,Kiani A,Shakiba Y,Rahimi Z,Bahrehmand F,Shakiba E,Vaisi-Raygani AA,Alibakhshi R,Eivazi A,Pourmotabbed T

    更新日期：2019-02-01 00:00:00

  • Genetic variants in transforming growth factor-β gene (TGFB1) affect susceptibility to schizophrenia.

    abstract：:Immense body of evidence indicates that dysfunction of immune system is implicated in the etiology of schizophrenia. The immune theory of schizophrenia is supported by alterations in cytokine profile in the brain and peripheral blood. Given the strong genetic background of schizophrenia, it might be assumed that aberr...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-013-2662-8

    authors： Frydecka D,Misiak B,Beszlej JA,Karabon L,Pawlak-Adamska E,Tomkiewicz A,Partyka A,Jonkisz A,Kiejna A

    更新日期：2013-10-01 00:00:00

  • 3-Deoxyglucosone interferes with insulin signaling and attenuates insulin action on glucose-induced GLP-1 secretion in the enteroendocrine L cell line STC-1.

    abstract：:Maintenance of glucose homeostasis is reciprocally regulated by insulin and glucagon-like peptide-1 (GLP-1). We previously reported that GLP-1 secretion in response to an oral glucose load was impaired following an administration of 3-deoxyglucosone (3DG), an independent factor associated with the development of pre-d...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-019-04926-0

    authors： Song X,Zhou L,Xu H,Wang F,Liang G,Zhang L,Huang F,Jiang G

    更新日期：2019-10-01 00:00:00

  • Haplotypes of CYP1B1 and CCDC57 genes in an Afro-Caribbean female population with uterine leiomyoma.

    abstract：:Uterine leiomyomas (UL) are prevalent benign tumors, especially among women of African ancestry. The disease also has genetic liability and is influenced by risk factors such as hormones and obesity. This study investigates the haplotypes of the Cytochrome P450 1B1 gene (CYP1B1) related to hormones and coiled-coil dom...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-019-04790-y

    authors： Alleyne AT,Bideau VS

    更新日期：2019-06-01 00:00:00

  • Cytotoxic and anti-angiogenic effects of zoledronic acid in DU-145 and PC-3 prostate cancer cell lines.

    abstract：:This study aims to investigate the apoptotic and anti-angiogenic effect of Zoledronic acid on hormone-refractory prostate cancer cell lines. XTT cell proliferation assay used to assess cytotoxicity. Caspase 3/7 activity and DNA fragmentation were measured to verify apoptosis. Angiogenic cytokine profiles investigated ...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-020-05840-6

    authors： Gokalp C

    更新日期：2020-10-01 00:00:00

  • The role of CcTpt1 in scale and early embryo development in common carp (Cyprinus carpio, Cyprinidae).

    abstract：:The full length cDNA sequence of the Tpt1/TCTP (Tumor protein, Translationally-controlled1) gene was identified from Common Carp (Cyprinus carpio, Cyprinidae), and was designated as CcTpt1 gene. The CDS is 510 bp and encodes a 170-amino acid peptide with a typical Tpt1 signature 2 domain, and is a typical Tpt1 protein...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-013-2806-x

    authors： Jiang L,Wang Y,Cheng A,Zhang B,Ma L,Liu Y,Sun X

    更新日期：2013-10-13 00:00:00

  • Development of bovine embryos in vitro in coculture with murine mesenchymal stem cells and embryonic fibroblasts.

    abstract：:Despite the progress on development of new culture media, in vitro-produced embryos still display lower quality when compared to the in vivo-produced counterparts. Coculture has been reconsidered as an alternative to improve embryo quality. Mesenchymal stem cells (MSC) and murine embryonic fibroblasts (MEF) have been ...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-018-4329-y

    authors： Ascari IJ,Martins SC,Camargo LSA,Mendez-Otero R,Jasmin

    更新日期：2018-12-01 00:00:00

  • Origanum vulgare essential oil: antibacterial activities and synergistic effect with polymyxin B against multidrug-resistant Acinetobacter baumannii.

    abstract：:Antimicrobial resistance is increasing around the world and the search for effective treatment options, such as new antibiotics and combination therapy is urgently needed. The present study evaluates oregano essential oil (OEO) antibacterial activities against reference and multidrug-resistant clinical isolates of Aci...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-020-05989-0

    authors： Amaral SC,Pruski BB,de Freitas SB,Allend SO,Ferreira MRA,Moreira C Jr,Pereira DIB,Junior ASV,Hartwig DD

    更新日期：2020-12-01 00:00:00

  • Molecular analysis of an actin gene, CarACT1, from chickpea (Cicer arietinum L.).

    abstract：:Actins are ubiquitous and highly conserved proteins that play key roles in cell formation and cellular activities. In this study, an actin gene was isolated from chickpea for the first time and designated as CarACT1 (for Cicer arietinum L. actin gene 1; Genbank accession no. EU529707). It encoded a putative protein wi...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-009-9844-4

    authors： Peng H,Cheng H,Yu X,Shi Q,Zhang H,Li J,Ma H

    更新日期：2010-02-01 00:00:00

  • Effects of the interleukin-1β-511 C/T gene polymorphism on the risk of gastric cancer in the context of the relationship between race and H. pylori infection: a meta-analysis of 20,000 subjects.

    abstract：:The interleukin (IL)-1β-511 C/T polymorphism has been shown to be functional and to contribute to the risk of gastric cancer. However, the relationship between the IL-1β-511 C/T polymorphism and gastric carcinogenesis remains inconclusive. A systematical electronic search was conducted of the MEDLINE, EMBASE, and CENT...

    journal_title：Molecular biology reports

    pub_type： 杂志文章,meta分析

    doi：10.1007/s11033-014-3748-7

    authors： Park MJ,Hyun MH,Yang JP,Yoon JM,Park S

    更新日期：2015-01-01 00:00:00

  • Ebselen, a promising antioxidant drug: mechanisms of action and targets of biological pathways.

    abstract：:Ebselen, an organoselenium compound, mimics glutathione peroxidase activity. It is a multifunctional compound, which catalyzes several essential reactions for the protection of cellular components from oxidative and free radical damage. Based on a number of in vitro and in vivo studies, various mechanisms are proposed...

    journal_title：Molecular biology reports

    pub_type： 杂志文章,评审

    doi：10.1007/s11033-014-3417-x

    authors： Azad GK,Tomar RS

    更新日期：2014-08-01 00:00:00

  • Innate endophytic fungus, Aspergillus terreus as biotic elicitor of withanolide A in root cell suspension cultures of Withania somnifera.

    abstract：:In the present study, root cell suspension cultures of W. somnifera were elicited with mycelial extract (1% w/v) and culture filtrate (5% v/v) of their native endophytic fungus Aspergillus terreus 2aWF in shake flask. Culture filtrate of A. terreus 2aWF significantly elicits withanolide A at 6H (12.20 ± 0.52 µg/g FCB)...

    journal_title：Molecular biology reports

    pub_type： 杂志文章

    doi：10.1007/s11033-019-04641-w

    authors： Kushwaha RK,Singh S,Pandey SS,Kalra A,Vivek Babu CS

    更新日期：2019-04-01 00:00:00