DOCSLIB.ORG

Mouse Aifm2 Knockout Project (CRISPR/Cas9)

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Mouse Aifm2 Knockout Project (CRISPR/Cas9) https://www.alphaknockout.com Mouse Aifm2 Knockout Project (CRISPR/Cas9) Objective: To create a Aifm2 knockout Mouse model (C57BL/6N) by CRISPR/Cas-mediated genome engineering. Strategy summary: The Aifm2 gene (NCBI Reference Sequence: NM_001039194 ; Ensembl: ENSMUSG00000020085 ) is located on Mouse chromosome 10. 9 exons are identified, with the ATG start codon in exon 2 and the TGA stop codon in exon 9 (Transcript: ENSMUST00000099706). Exon 4~8 will be selected as target site. Cas9 and gRNA will be co-injected into fertilized eggs for KO Mouse production. The pups will be genotyped by PCR followed by sequencing analysis. Note: Homozygous null mice display reduced sensitivity to genotoxin induced cellular growth inhibition but have no change in spontaneous or induced tumor incidence. Exon 4 starts from about 26.36% of the coding region. Exon 4~8 covers 60.41% of the coding region. The size of effective KO region: ~8012 bp. The KO region does not have any other known gene. Page 1 of 9 https://www.alphaknockout.com Overview of the Targeting Strategy Wildtype allele 5' gRNA region gRNA region 3' 1 4 5 6 7 8 9 Legends Exon of mouse Aifm2 Knockout region Page 2 of 9 https://www.alphaknockout.com Overview of the Dot Plot (up) Window size: 15 bp Forward Reverse Complement Sequence 12 Note: The 1166 bp section upstream of Exon 4 is aligned with itself to determine if there are tandem repeats. No significant tandem repeat is found in the dot plot matrix. So this region is suitable for PCR screening or sequencing analysis. Overview of the Dot Plot (down) Window size: 15 bp Forward Reverse Complement Sequence 12 Note: The 924 bp section downstream of Exon 8 is aligned with itself to determine if there are tandem repeats. Tandem repeats are found in the dot plot matrix. The gRNA site is selected outside of these tandem repeats. Page 3 of 9 https://www.alphaknockout.com Overview of the GC Content Distribution (up) Window size: 300 bp Sequence 12 Summary: Full Length(1166bp) | A(21.78% 254) | C(24.87% 290) | T(28.9% 337) | G(24.44% 285) Note: The 1166 bp section upstream of Exon 4 is analyzed to determine the GC content. No significant high GC-content region is found. So this region is suitable for PCR screening or sequencing analysis. Overview of the GC Content Distribution (down) Window size: 300 bp Sequence 12 Summary: Full Length(924bp) | A(31.06% 287) | C(23.7% 219) | T(23.81% 220) | G(21.43% 198) Note: The 924 bp section downstream of Exon 8 is analyzed to determine the GC content. No significant high GC-content region is found. So this region is suitable for PCR screening or sequencing analysis. Page 4 of 9 https://www.alphaknockout.com BLAT Search Results (up) QUERY SCORE START END QSIZE IDENTITY CHROM STRAND START END SPAN ----------------------------------------------------------------------------------------------- browser details YourSeq 1166 1 1166 1166 100.0% chr10 + 61726482 61727647 1166 browser details YourSeq 309 446 1138 1166 94.1% chr11 - 70310901 70556394 245494 browser details YourSeq 213 517 1137 1166 87.0% chr11 - 74709871 74710450 580 browser details YourSeq 211 513 1140 1166 84.6% chr4 + 123755405 123755838 434 browser details YourSeq 202 513 1144 1166 84.4% chr13 - 55288730 55289069 340 browser details YourSeq 188 546 1151 1166 95.7% chr4 + 133564756 133565371 616 browser details YourSeq 180 529 1137 1166 86.3% chr1 + 160922368 160922886 519 browser details YourSeq 171 933 1137 1166 94.3% chr1 + 88603387 88603591 205 browser details YourSeq 169 946 1140 1166 91.5% chr18 - 76812964 76813152 189 browser details YourSeq 168 940 1138 1166 95.2% chr4 - 40250567 40250765 199 browser details YourSeq 167 952 1155 1166 92.5% chr11 - 6609565 6609763 199 browser details YourSeq 167 615 1137 1166 85.7% chr11 + 23259836 23260036 201 browser details YourSeq 166 946 1154 1166 89.6% chr2 + 177532069 177532269 201 browser details YourSeq 166 946 1139 1166 94.6% chr11 + 88519804 88519996 193 browser details YourSeq 165 699 1138 1166 87.2% chr8 - 125661881 125662090 210 browser details YourSeq 163 952 1144 1166 95.6% chr13 - 74904202 74904395 194 browser details YourSeq 162 944 1139 1166 94.6% chr9 - 78067966 78068164 199 browser details YourSeq 162 952 1156 1166 91.3% chr9 - 72670784 72670984 201 browser details YourSeq 162 952 1140 1166 92.7% chr6 - 49447535 49447717 183 browser details YourSeq 161 622 1140 1166 85.5% chr2 - 32240168 32240358 191 Note: The 1166 bp section upstream of Exon 4 is BLAT searched against the genome. No significant similarity is found. BLAT Search Results (down) QUERY SCORE START END QSIZE IDENTITY CHROM STRAND START END SPAN ----------------------------------------------------------------------------------------------- browser details YourSeq 924 1 924 924 100.0% chr10 + 61735660 61736583 924 browser details YourSeq 136 136 313 924 92.1% chr10 + 117064448 117064957 510 browser details YourSeq 132 118 313 924 89.8% chr13 + 14046481 14046681 201 browser details YourSeq 130 134 305 924 89.7% chr13 + 104502258 104502445 188 browser details YourSeq 128 132 302 924 88.6% chr13 + 52493433 52493618 186 browser details YourSeq 127 134 298 924 93.3% chr10 + 61161469 61162044 576 browser details YourSeq 125 134 307 924 91.0% chr13 + 62431993 62432179 187 browser details YourSeq 123 133 298 924 89.2% chr1 + 186771755 186771936 182 browser details YourSeq 117 134 303 924 90.5% chr14 + 65141016 65141188 173 browser details YourSeq 116 156 311 924 93.4% chr13 + 64143690 64143859 170 browser details YourSeq 115 156 297 924 92.0% chr2 - 154068765 154068922 158 browser details YourSeq 115 134 297 924 90.8% chr10 + 122923478 122923652 175 browser details YourSeq 114 157 305 924 92.6% chr3 - 58502976 58503147 172 browser details YourSeq 113 156 298 924 91.8% chr11 - 86775011 86775163 153 browser details YourSeq 112 134 305 924 89.2% chr7 - 27959917 27960087 171 browser details YourSeq 111 149 295 924 89.6% chr11 - 84128298 84128461 164 browser details YourSeq 111 166 298 924 93.1% chr8 + 119772862 119773005 144 browser details YourSeq 107 149 304 924 92.3% chr13 - 54521605 54521798 194 browser details YourSeq 107 149 656 924 94.2% chr7 + 80800632 80801237 606 browser details YourSeq 107 133 289 924 92.2% chr4 + 133126741 133126904 164 Note: The 924 bp section downstream of Exon 8 is BLAT searched against the genome. No significant similarity is found. Page 5 of 9 https://www.alphaknockout.com Gene and protein information: Aifm2 apoptosis-inducing factor, mitochondrion-associated 2 [ Mus musculus (house mouse) ] Gene ID: 71361, updated on 12-Aug-2019 Gene summary Official Symbol Aifm2 provided by MGI Official Full Name apoptosis-inducing factor, mitochondrion-associated 2 provided by MGI Primary source MGI:MGI:1918611 See related Ensembl:ENSMUSG00000020085 Gene type protein coding RefSeq status VALIDATED Organism Mus musculus Lineage Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae; Murinae; Mus; Mus Also known as Amid; PRG3; 5430437E11Rik; D730001I10Rik Expression Biased expression in adrenal adult (RPKM 65.8), bladder adult (RPKM 44.5) and 13 other tissues See more Orthologs human all Genomic context Location: 10; 10 B4 See Aifm2 in Genome Data Viewer Exon count: 14 Annotation release Status Assembly Chr Location 108 current GRCm38.p6 (GCF_000001635.26) 10 NC_000076.6 (61715263..61739260) Build 37.2 previous assembly MGSCv37 (GCF_000001635.18) 10 NC_000076.5 (61178011..61202008) Chromosome 10 - NC_000076.6 Page 6 of 9 https://www.alphaknockout.com Transcript information: This gene has 7 transcripts Gene: Aifm2 ENSMUSG00000020085 Description apoptosis-inducing factor, mitochondrion-associated 2 [Source:MGI Symbol;Acc:MGI:1918611] Gene Synonyms 5430437E11Rik, Amid, D730001I10Rik, PRG3 Location Chromosome 10: 61,715,263-61,739,260 forward strand. GRCm38:CM001003.2 About this gene This gene has 7 transcripts (splice variants), 204 orthologues, is a member of 1 Ensembl protein family and is associated with 1 phenotype. Transcripts Name Transcript ID bp Protein Translation ID Biotype CCDS UniProt Flags Aifm2-203 ENSMUST00000099706.8 3086 373aa ENSMUSP00000097297.2 Protein coding CCDS23884 Q8BUE4 TSL:1 GENCODE basic APPRIS P1 Aifm2-204 ENSMUST00000105455.8 1512 373aa ENSMUSP00000101095.2 Protein coding CCDS23884 Q8BUE4 TSL:1 GENCODE basic APPRIS P1 Aifm2-201 ENSMUST00000067857.10 1442 373aa ENSMUSP00000070054.4 Protein coding CCDS23884 Q8BUE4 TSL:1 GENCODE basic APPRIS P1 Aifm2-202 ENSMUST00000080099.5 1329 380aa ENSMUSP00000078998.5 Protein coding CCDS48576 Q8BUE4 TSL:1 GENCODE basic Aifm2-206 ENSMUST00000131361.1 656 No protein - Retained intron - - TSL:2 Aifm2-207 ENSMUST00000140664.1 477 No protein - Retained intron - - TSL:3 Aifm2-205 ENSMUST00000129809.1 378 No protein - lncRNA - - TSL:5 Page 7 of 9 https://www.alphaknockout.com 44.00 kb Forward strand 61.71Mb 61.72Mb 61.73Mb 61.74Mb Genes (Comprehensive set... Aifm2-204 >protein coding Aifm2-206 >retained intron Aifm2-207 >retained intron Aifm2-201 >protein coding Aifm2-203 >protein coding Aifm2-205 >lncRNA Aifm2-202 >protein coding Contigs AC153136.3 > Genes < H2afy2-201protein coding (Comprehensive set... Regulatory Build 61.71Mb 61.72Mb 61.73Mb 61.74Mb Reverse strand 44.00 kb Regulation Legend CTCF Enhancer Open Chromatin Promoter Promoter Flank Transcription Factor Binding Site Gene Legend Protein Coding merged Ensembl/Havana Ensembl protein coding Non-Protein Coding RNA gene processed transcript Page 8 of 9 https://www.alphaknockout.com Transcript: ENSMUST00000099706 23.32 kb Forward strand Aifm2-203 >protein coding ENSMUSP00000097... Transmembrane heli... Low complexity (Seg) Superfamily FAD/NAD(P)-binding domain superfamily Prints PR00368 PR00469 Pfam FAD/NAD(P)-binding domain PANTHER PTHR43735:SF3 PTHR43735 Gene3D 3.50.50.100 All sequence SNPs/i... Sequence variants (dbSNP and all other sources) Variant Legend missense variant splice region variant synonymous variant Scale bar 0 40 80 120 160 200 240 280 320 373 We wish to acknowledge the following valuable scientific information resources: Ensembl, MGI, NCBI, UCSC.
Load more

Recommended publications
  • AIFM2 Monoclonal Antibody (M13A), Clone 2C6
    AIFM2 monoclonal antibody (M13A), clone 2C6 Catalog # : H00084883-M13A 規格 : [ 200 uL ] List All Specification Application Image Product Mouse monoclonal antibody raised against a partial recombinant Western Blot (Transfected lysate) Description: AIFM2. Immunogen: AIFM2 (AAH06121, 1 a.a. ~ 339 a.a) partial recombinant protein with GST tag. MW of the GST tag alone is 26 KDa. Sequence: MGSQVSVESGALHVVIVGGGFGGIAAASQLQALNVPFMLVDMKDSFHHN VAALRASVETGFAKKTFISYSVTFKDNFRQGLVVGIDLKNQMVLLQGGE ALPFSHLILATGSTGPFPGKFNEVSSQQAAIQAYEDMVRQVQRSRFIVVV enlarge GGGSAGVEMAAEIKTEYPEKEVTLIHSQVALADKELLPSVRQEVKEILLRK Western Blot (Recombinant GVQLLLSERVSNLEELPLNEYREYIKVQTDKGTEVATNLVILCTGIKINSSA protein) YRKAFESRLASSGALRVNEHLQVEGHSNVYAIGDCADVRTPKMAYLAGL HANIAVANIVNSVKQRPLQAYKPGALTFLLSMGRNDGVG ELISA Host: Mouse Reactivity: Human Isotype: IgG1 Kappa Quality Control Antibody Reactive Against Recombinant Protein. Testing: Western Blot detection against Immunogen (62.92 KDa) . Storage Buffer: In ascites fluid Storage Store at -20°C or lower. Aliquot to avoid repeated freezing and thawing. Instruction: MSDS: Download Interspecies Mouse (90); Rat (90) Antigen Sequence: Datasheet: Download Applications Western Blot (Transfected lysate) Page 1 of 3 2021/6/18 Western Blot analysis of AIFM2 expression in transfected 293T cell line by AMID monoclonal antibody (M13A), clone 2C6. Lane 1: AIFM2 transfected lysate(40.5 KDa). Lane 2: Non-transfected lysate. Protocol Download Western Blot (Recombinant protein) Protocol Download ELISA Gene Information Entrez GeneID: 84883 GeneBank BC006121 Accession#: Protein AAH06121 Accession#: Gene Name: AIFM2 Gene Alias: AMID,PRG3,RP11-367H5.2 Gene apoptosis-inducing factor, mitochondrion-associated, 2 Description: Omim ID: 605159 Gene Ontology: Hyperlink Gene Summary: The protein encoded by this gene has significant homology to NADH oxidoreductases and the apoptosis-inducing factor PDCD8/AIF. Overexpression of this gene has been shown to induce apoptosis. The expression of this gene is found to be induced by tumor suppressor protein p53 in colon caner cells.
    [Show full text]
  • UNIVERSITY of CALIFORNIA RIVERSIDE Investigations Into The
    UNIVERSITY OF CALIFORNIA RIVERSIDE Investigations into the Role of TAF1-mediated Phosphorylation in Gene Regulation A Dissertation submitted in partial satisfaction of the requirements for the degree of Doctor of Philosophy in Cell, Molecular and Developmental Biology by Brian James Gadd December 2012 Dissertation Committee: Dr. Xuan Liu, Chairperson Dr. Frank Sauer Dr. Frances M. Sladek Copyright by Brian James Gadd 2012 The Dissertation of Brian James Gadd is approved Committee Chairperson University of California, Riverside Acknowledgments I am thankful to Dr. Liu for her patience and support over the last eight years. I am deeply indebted to my committee members, Dr. Frank Sauer and Dr. Frances Sladek for the insightful comments on my research and this dissertation. Thanks goes out to CMDB, especially Dr. Bachant, Dr. Springer and Kathy Redd for their support. Thanks to all the members of the Liu lab both past and present. A very special thanks to the members of the Sauer lab, including Silvia, Stephane, David, Matt, Stephen, Ninuo, Toby, Josh, Alice, Alex and Flora. You have made all the years here fly by and made them so enjoyable. From the Sladek lab I want to thank Eugene, John, Linh and Karthi. Special thanks go out to all the friends I’ve made over the years here. Chris, Amber, Stephane and David, thank you so much for feeding me, encouraging me and keeping me sane. Thanks to the brothers for all your encouragement and prayers. To any I haven’t mentioned by name, I promise I haven’t forgotten all you’ve done for me during my graduate years.
    [Show full text]
  • Integrative Analysis of Genomic Amplification-Dependent Expression
    Oncogene (2020) 39:4118–4131 https://doi.org/10.1038/s41388-020-1279-3 ARTICLE Integrative analysis of genomic amplification-dependent expression and loss-of-function screen identifies ASAP1 as a driver gene in triple-negative breast cancer progression 1 1 1 2 2 Jichao He ● Ronan P. McLaughlin ● Lambert van der Beek ● Sander Canisius ● Lodewyk Wessels ● 3 3 3 1 1 Marcel Smid ● John W. M. Martens ● John A. Foekens ● Yinghui Zhang ● Bob van de Water Received: 26 September 2019 / Revised: 14 March 2020 / Accepted: 17 March 2020 / Published online: 31 March 2020 © The Author(s) 2020. This article is published with open access Abstract The genetically heterogeneous triple-negative breast cancer (TNBC) continues to be an intractable disease, due to lack of effective targeted therapies. Gene amplification is a major event in tumorigenesis. Genes with amplification-dependent expression are being explored as therapeutic targets for cancer treatment. In this study, we have applied Analytical Multi- scale Identification of Recurring Events analysis and transcript quantification in the TNBC genome across 222 TNBC tumors and identified 138 candidate genes with positive correlation in copy number gain (CNG) and gene expression. siRNA-based 1234567890();,: 1234567890();,: loss-of-function screen of the candidate genes has validated EGFR, MYC, ASAP1, IRF2BP2, and CCT5 genes as drivers promoting proliferation in different TNBC cells. MYC, ASAP1, IRF2BP2, and CCT5 display frequent CNG and concurrent expression over 2173 breast cancer tumors (cBioPortal dataset). More frequently are MYC and ASAP1 amplified in TNBC tumors (>30%, n = 320). In particular, high expression of ASAP1, the ADP-ribosylation factor GTPase-activating protein, is significantly related to poor metastatic relapse-free survival of TNBC patients (n = 257, bc-GenExMiner).
    [Show full text]
  • A Dissertation Submitted to the Faculty of the Graduate School of Arts And
    MOLECULAR MODULATION OF ESTROGEN -INDUCED APOPTOSIS IN LONG -TERM ESTROGEN - DEPRIVED BREAST CANCER CELLS A Dissertation submitted to the Faculty of the Graduate School of Arts and Sciences of Georgetown University in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Tumor Biology By Elizabeth E. Sweeney, B.S. Washington, DC March 31, 2014 Copyright 2014 by Elizabeth E. Sweeney All Rights Reserved ii MOLECULAR MODULATION OF ESTROGEN -INDUCED APOPTOSIS IN LONG -TERM ESTROGEN - DEPRIVED BREAST CANCER CELLS Elizabeth E. Sweeney, B.S. Thesis Advisor: V. Craig Jordan , O.B.E., Ph.D., D.Sc., F.Med.Sci . ABSTRACT Estrogen receptor (ER)-positive breast cancer cell lines have been instrumental in modeling breast cancer and providing an opportunity to document the development and evolution of acquired anti-hormone resistance. Models of long-term estrogen-deprived breast cancer cells are utilized in the laboratory to mimic clinical aromatase inhibitor-resistant breast cancer and serve as a tool to discover new therapeutic strategies. The MCF-7:5C and MCF-7:2A subclones were generated through long-term estrogen deprivation of ER-positive MCF-7 cells, and represent anti-hormone resistant breast cancer. MCF-7:5C cells paradoxically undergo estrogen-induced apoptosis within seven days of estrogen (estradiol, E 2) treatment; MCF-7:2A cells also experience E2-induced apoptosis but evade dramatic cell death until approximately 14 days of treatment. Our data suggest that MCF-7:2A cells employ stronger antioxidant defense mechanisms than do MCF-7:5C cells, and that oxidative stress is ultimately required for MCF- 7:2A cells to die in response to E2 treatment.
    [Show full text]
  • Ferroptosis-Related Flavoproteins: Their Function and Stability
    International Journal of Molecular Sciences Review Ferroptosis-Related Flavoproteins: Their Function and Stability R. Martin Vabulas Charité-Universitätsmedizin, Institute of Biochemistry, Charitéplatz 1, 10117 Berlin, Germany; [email protected]; Tel.: +49-30-4505-28176 Abstract: Ferroptosis has been described recently as an iron-dependent cell death driven by peroxida- tion of membrane lipids. It is involved in the pathogenesis of a number of diverse diseases. From the other side, the induction of ferroptosis can be used to kill tumor cells as a novel therapeutic approach. Because of the broad clinical relevance, a comprehensive understanding of the ferroptosis-controlling protein network is necessary. Noteworthy, several proteins from this network are flavoenzymes. This review is an attempt to present the ferroptosis-related flavoproteins in light of their involvement in anti-ferroptotic and pro-ferroptotic roles. When available, the data on the structural stability of mutants and cofactor-free apoenzymes are discussed. The stability of the flavoproteins could be an important component of the cellular death processes. Keywords: flavoproteins; riboflavin; ferroptosis; lipid peroxidation; protein quality control 1. Introduction Human flavoproteome encompasses slightly more than one hundred enzymes that par- ticipate in a number of key metabolic pathways. The chemical versatility of flavoproteins relies on the associated cofactors, flavin mononucleotide (FMN) and flavin adenine dinu- cleotide (FAD). In humans, flavin cofactors are biosynthesized from a precursor riboflavin that has to be supplied with food. To underline its nutritional essentiality, riboflavin is called vitamin B2. In compliance with manifold cellular demands, flavoproteins have been accommo- Citation: Vabulas, R.M. dated to operate at different subcellular locations [1].
    [Show full text]
  • Theranostics ATF6 Aggravates Acinar Cell Apoptosis and Injury By
    Theranostics 2020, Vol. 10, Issue 18 8298 Ivyspring International Publisher Theranostics 2020; 10(18): 8298-8314. doi: 10.7150/thno.46934 Research Paper ATF6 aggravates acinar cell apoptosis and injury by regulating p53/AIFM2 transcription in Severe Acute Pancreatitis Jie-Hui Tan1#, Rong-Chang Cao1#, Lei Zhou1, Zhi-Tao Zhou2, Huo-Ji Chen3, Jia Xu4, Xue-Mei Chen5, Yang-Chen Jin6, Jia-Yu Lin6, Jun-Ling Zeng7, Shu-Ji Li8, Min Luo9, Guo-Dong Hu10, Xiao-Bing Yang11, Jin Jin12 and Guo-Wei Zhang1 1. Division of Hepatobiliopancreatic Surgery, Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, China. 2. Department of the Electronic Microscope Room, Central Laboratory, Southern Medical University, Guangzhou, China. 3. School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, China. 4. Department of Pathophysiology, Southern Medical University, Guangzhou, China. 5. Department of Occupational Health and Medicine, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, China. 6. The First Clinical Medical College, Southern Medical University, Guangzhou, China. 7. Laboratory Animal Research Center of Nanfang Hospital, Southern Medical University, Guangzhou, China. 8. Guangdong-Hong Kong-Macao Greater Bay Area Center for Brain Science and Brain-Inspired Intelligence, Southern Medical University, Guangzhou, China. 9. Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, China. 10. Department of Respiratory and Crit Care Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, China. 11. Division of Nephrology, Nanfang Hospital, Southern Medical University, National Clinical Research Center for Kidney Disease, State Key Laboratory of Organ Failure Research, Guangdong Institute, Guangzhou, China. 12. Department of Gynaecology and Obstetrics, Nanfang Hospital, Southern Medical University, Guangzhou, China.
    [Show full text]
  • Identification of Genomic Targets of Krüppel-Like Factor 9 in Mouse Hippocampal
    Identification of Genomic Targets of Krüppel-like Factor 9 in Mouse Hippocampal Neurons: Evidence for a role in modulating peripheral circadian clocks by Joseph R. Knoedler A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Neuroscience) in the University of Michigan 2016 Doctoral Committee: Professor Robert J. Denver, Chair Professor Daniel Goldman Professor Diane Robins Professor Audrey Seasholtz Associate Professor Bing Ye ©Joseph R. Knoedler All Rights Reserved 2016 To my parents, who never once questioned my decision to become the other kind of doctor, And to Lucy, who has pushed me to be a better person from day one. ii Acknowledgements I have a huge number of people to thank for having made it to this point, so in no particular order: -I would like to thank my adviser, Dr. Robert J. Denver, for his guidance, encouragement, and patience over the last seven years; his mentorship has been indispensable for my growth as a scientist -I would also like to thank my committee members, Drs. Audrey Seasholtz, Dan Goldman, Diane Robins and Bing Ye, for their constructive feedback and their willingness to meet in a frequently cold, windowless room across campus from where they work -I am hugely indebted to Pia Bagamasbad and Yasuhiro Kyono for teaching me almost everything I know about molecular biology and bioinformatics, and to Arasakumar Subramani for his tireless work during the home stretch to my dissertation -I am grateful for the Neuroscience Program leadership and staff, in particular
    [Show full text]
  • Novel Role of IMPA2 in AIFM2-Mediated Apoptosis of Cervical Cancer by Targeting P53
    Novel Role of IMPA2 in AIFM2-Mediated Apoptosis of Cervical Cancer by Targeting p53 Lei Liu Second Xiangya Hospital Min Wang ( [email protected] ) Second Xiangya Hospital https://orcid.org/0000-0002-3420-4031 Xianping Li Second Xiangya Hospital Bingqi Wang Second Xiangya Hospital Sheng Yin Second Xiangya Hospital Primary research Keywords: IMPA2, apoptosis, AIFM2, p53, cervical cancer Posted Date: March 4th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-263816/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/22 Abstract Background Cervical cancer continues to be concerned and the prognosis of locally advanced cervical cancer remains poor, which underscores pivotal needs to nd novel therapeutic targets. Previously, we rstly identied Myo-inositol monophosphatase 2(IMPA2) as a potential oncogene and veried its tumor-promoting role in vitro and in vivo. In this study, we further aimed to elucidate the underlying mechanisms of IMPA2 in regulation of tumor apoptosis. Methods Cell apoptosis was assessed by apoptosis-related proteins detecting, ow cytometry, immunouorescence and immunohistochemical staining. Fluorescence microscope was used to analyze uorescence signal. The string database was used to nd molecules regulated by IMPA2. The expression of apoptosis inducing factor mitochondria associated 2(AIFM2) was determined by qRT-PCR, western blot, immunohistochemical staining and immunouorescence analysis. Function changes of mitochondria were evaluated by measurements of mitochondrial membrane potential and intracellular Ca2+ levels. CCK-8 assay was used to detected cell viability. Results Apoptosis of cervical cancer cells was markedly promoted when silencing IMPA2. AIFM2 was signicantly up-regulated both in mRNA and protein levels, and inhibition of AIFM2 could reserve IMPA2 knockdown-induced apoptosis in a mitochondrial dependent manner.
    [Show full text]
  • Arginine Methyltransferase PRMT1 Regulates P53 Activity in Breast Cancer
    life Article Arginine Methyltransferase PRMT1 Regulates p53 Activity in Breast Cancer Li-Ming Liu 1,2,3,†, Qiang Tang 1,2,3,†, Xin Hu 1,2,3,4, Jing-Jing Zhao 1,2, Yuan Zhang 5, Guo-Guang Ying 1,2,3,* and Fei Zhang 1,2,3,4,* 1 Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, China; [email protected] (L.-M.L.); [email protected] (Q.T.); [email protected] (X.H.); [email protected] (J.-J.Z.) 2 National Clinical Research Center for Cancer, Tianjin 300060, China 3 Tianjin’s Clinical Research Center for Cancer, Tianjin 300060, China 4 Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060, China 5 Department of International Medical Services, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100005, China; [email protected] * Correspondence: [email protected] (G.-G.Y.); [email protected] (F.Z.) † These authors contributed equally to this article. Abstract: The protein p53 is one of the most important tumor suppressors, responding to a variety of stress signals. Mutations in p53 occur in about half of human cancer cases, and dysregulation of the p53 function by epigenetic modifiers and modifications is prevalent in a large proportion of the re- mainder. PRMT1 is the main enzyme responsible for the generation of asymmetric-dimethylarginine, whose upregulation or aberrant splicing has been observed in many types of malignancies. Here, we demonstrate that p53 function is regulated by PRMT1 in breast cancer cells. PRMT1 knockdown activated the p53 signal pathway and induced cell growth-arrest and senescence.
    [Show full text]
  • Ferroptosis: Past, Present and Future Jie Li 1,2,Fengcao3, He-Liang Yin4,5, Zi-Jian Huang1,2, Zhi-Tao Lin1,2,Ningmao1,2,Beisun1,2 and Gang Wang1,2
    Li et al. Cell Death and Disease (2020) 11:88 https://doi.org/10.1038/s41419-020-2298-2 Cell Death & Disease REVIEW ARTICLE Open Access Ferroptosis: past, present and future Jie Li 1,2,FengCao3, He-liang Yin4,5, Zi-jian Huang1,2, Zhi-tao Lin1,2,NingMao1,2,BeiSun1,2 and Gang Wang1,2 Abstract Ferroptosis is a new type of cell death that was discovered in recent years and is usually accompanied by a large amount of iron accumulation and lipid peroxidation during the cell death process; the occurrence of ferroptosis is iron-dependent. Ferroptosis-inducing factors can directly or indirectly affect glutathione peroxidase through different pathways, resulting in a decrease in antioxidant capacity and accumulation of lipid reactive oxygen species (ROS) in cells, ultimately leading to oxidative cell death. Recent studies have shown that ferroptosis is closely related to the pathophysiological processes of many diseases, such as tumors, nervous system diseases, ischemia-reperfusion injury, kidney injury, and blood diseases. How to intervene in the occurrence and development of related diseases by regulating cell ferroptosis has become a hotspot and focus of etiological research and treatment, but the functional changes and specific molecular mechanisms of ferroptosis still need to be further explored. This paper systematically summarizes the latest progress in ferroptosis research, with a focus on providing references for further understanding of its pathogenesis and for proposing new targets for the treatment of related diseases. Facts peroxides, or can other substances take the place of iron in ferroptosis? What is the downstream regulation mechanism of iron 1234567890():,; 1234567890():,; 1234567890():,; 1234567890():,; Ferroptosis is a new type of programmed cell death, in ferroptosis? which occurs with iron dependence.
    [Show full text]
  • Non-Apoptotic Cell Death Signaling Pathways in Melanoma
    International Journal of Molecular Sciences Review Non-Apoptotic Cell Death Signaling Pathways in Melanoma Mariusz L. Hartman Department of Molecular Biology of Cancer, Medical University of Lodz, 6/8 Mazowiecka Street, 92-215 Lodz, Poland; [email protected]; Tel.: +48-42-272-57-03 Received: 10 April 2020; Accepted: 22 April 2020; Published: 23 April 2020 Abstract: Resisting cell death is a hallmark of cancer. Disturbances in the execution of cell death programs promote carcinogenesis and survival of cancer cells under unfavorable conditions, including exposition to anti-cancer therapies. Specific modalities of regulated cell death (RCD) have been classified based on different criteria, including morphological features, biochemical alterations and immunological consequences. Although melanoma cells are broadly equipped with the anti-apoptotic machinery and recurrent genetic alterations in the components of the RAS/RAF/MEK/ERK signaling markedly contribute to the pro-survival phenotype of melanoma, the roles of autophagy-dependent cell death, necroptosis, ferroptosis, pyroptosis, and parthanatos have recently gained great interest. These signaling cascades are involved in melanoma cell response and resistance to the therapeutics used in the clinic, including inhibitors of BRAFmut and MEK1/2, and immunotherapy. In addition, the relationships between sensitivity to non-apoptotic cell death routes and specific cell phenotypes have been demonstrated, suggesting that plasticity of melanoma cells can be exploited to modulate response of these cells to different cell death stimuli. In this review, the current knowledge on the non-apoptotic cell death signaling pathways in melanoma cell biology and response to anti-cancer drugs has been discussed. Keywords: autophagy; differentiation; drug resistance; ferroptosis; melanoma; necroptosis; parthanatos; pyroptosis; reactive oxygen species (ROS); targeted therapy 1.
    [Show full text]
  • Autocrine IFN Signaling Inducing Profibrotic Fibroblast Responses By
    Downloaded from http://www.jimmunol.org/ by guest on September 23, 2021 Inducing is online at: average * The Journal of Immunology , 11 of which you can access for free at: 2013; 191:2956-2966; Prepublished online 16 from submission to initial decision 4 weeks from acceptance to publication August 2013; doi: 10.4049/jimmunol.1300376 http://www.jimmunol.org/content/191/6/2956 A Synthetic TLR3 Ligand Mitigates Profibrotic Fibroblast Responses by Autocrine IFN Signaling Feng Fang, Kohtaro Ooka, Xiaoyong Sun, Ruchi Shah, Swati Bhattacharyya, Jun Wei and John Varga J Immunol cites 49 articles Submit online. Every submission reviewed by practicing scientists ? is published twice each month by Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts http://jimmunol.org/subscription Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html http://www.jimmunol.org/content/suppl/2013/08/20/jimmunol.130037 6.DC1 This article http://www.jimmunol.org/content/191/6/2956.full#ref-list-1 Information about subscribing to The JI No Triage! Fast Publication! Rapid Reviews! 30 days* Why • • • Material References Permissions Email Alerts Subscription Supplementary The Journal of Immunology The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2013 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. This information is current as of September 23, 2021. The Journal of Immunology A Synthetic TLR3 Ligand Mitigates Profibrotic Fibroblast Responses by Inducing Autocrine IFN Signaling Feng Fang,* Kohtaro Ooka,* Xiaoyong Sun,† Ruchi Shah,* Swati Bhattacharyya,* Jun Wei,* and John Varga* Activation of TLR3 by exogenous microbial ligands or endogenous injury-associated ligands leads to production of type I IFN.
    [Show full text]