* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download Bioteh_Klonesana un in vivo inhenierija_2015
Genetic code wikipedia , lookup
DNA profiling wikipedia , lookup
Comparative genomic hybridization wikipedia , lookup
DNA polymerase wikipedia , lookup
Nutriepigenomics wikipedia , lookup
Designer baby wikipedia , lookup
Genetic engineering wikipedia , lookup
Pathogenomics wikipedia , lookup
Transposable element wikipedia , lookup
Cancer epigenetics wikipedia , lookup
Minimal genome wikipedia , lookup
Mitochondrial DNA wikipedia , lookup
Microevolution wikipedia , lookup
DNA damage theory of aging wikipedia , lookup
SNP genotyping wikipedia , lookup
Gel electrophoresis of nucleic acids wikipedia , lookup
DNA vaccination wikipedia , lookup
Vectors in gene therapy wikipedia , lookup
United Kingdom National DNA Database wikipedia , lookup
Molecular cloning wikipedia , lookup
Genealogical DNA test wikipedia , lookup
DNA sequencing wikipedia , lookup
Site-specific recombinase technology wikipedia , lookup
Primary transcript wikipedia , lookup
DNA supercoil wikipedia , lookup
Nucleic acid double helix wikipedia , lookup
Cell-free fetal DNA wikipedia , lookup
Epigenomics wikipedia , lookup
Cre-Lox recombination wikipedia , lookup
Point mutation wikipedia , lookup
Extrachromosomal DNA wikipedia , lookup
Genome evolution wikipedia , lookup
Bisulfite sequencing wikipedia , lookup
Human genome wikipedia , lookup
Human Genome Project wikipedia , lookup
Microsatellite wikipedia , lookup
History of genetic engineering wikipedia , lookup
Whole genome sequencing wikipedia , lookup
Nucleic acid analogue wikipedia , lookup
Genomic library wikipedia , lookup
Zinc finger nuclease wikipedia , lookup
Metagenomics wikipedia , lookup
Non-coding DNA wikipedia , lookup
Artificial gene synthesis wikipedia , lookup
Deoxyribozyme wikipedia , lookup
No-SCAR (Scarless Cas9 Assisted Recombineering) Genome Editing wikipedia , lookup
Helitron (biology) wikipedia , lookup
BIOTEHNOLOĢIJA III : REKOMBINANTU BIOTEHNOLOĢIJA JAUNĀ BIOTEHNOLOĢIJA I. Muižnieks, 2015. g. pavasaris Jaunās klonēšanas un in vivo gēnu inženierijas metodes Klonēšanas metodes: Gibsona savākšanās In vivo gēnu inženierijas metodes: Zn-pirkstu nukleāzes TALEN nukleāzes CRISP/Cas sistēma Sekvenēšanas metožu attīstība W-H Chen, Z-J Qin, J Wang,G-P Zhao. The MASTER (methylation-assisted tailorable ends rational) ligation method for seamless DNA assembly. Nucleic Acids Research, 2013, 1–9, doi:10.1093/nar/gkt122 H. Kim & J-S Kim. A guide to genome engineering with programmable nucleases Nature Reviews Genetics 15, 321–334, (2014) doi:10.1038/nrg3686 a | Nuclease-induced double-strand breaks (DSBs) can lead to sequence insertion, nucleotide correction or change (red box) through homology-directed repair (HDR) in the presence of a donor DNA or a single-strand oligodeoxynucleotide (ssODN), both of which contain homology arms. DSBs can also be repaired through error-prone non-homologous end-joining (NHEJ), which does not require donor DNA or ssODN and consequently often leads to small insertions and deletions (indels). Typical indel sequences and the number of inserted (+3 and +1) or deleted (−2, −4 and −10) bases are shown. b | When two DSBs are generated in cis on a single chromosome by programmable nucleases, the flanking region can be deleted or inverted. c | When two DSBs are generated on two different chromosomes, chromosomal translocations can be induced. Jaunās pieejas augu genoma modifikācijai Zn-pirkstu nukleāzes Genes VII, Lewin B., 2005, p. 276/382 Jaunās pieejas augu genoma modifikācijai Zn-pirkstu nukleāzes Modificēta FokI endonukleāze Zinc-finger nucleases as gene therapy agents, D Carroll, 2008 A schematic representation of a zinc-finger nuclease (ZFN) pair a | Each ZFN is composed of a zinc-finger protein (ZFP) at the amino terminus and the FokI nuclease domain at the carboxyl terminus. In the zinc-finger motif consensus, X represents any amino acid. Target sequences of ZFN pairs are typically 18–36 bp in length, excluding spacers. b | A computer model structure of a ZFN pair bound to DNA is shown. Each zinc-finger is shown in shades of pink in ribbon (left) and space-filling (right) representations. The grey region represents the linker between the DNA-binding and catalytic domains. The FokI catalytic domains are shown in blue and purple at the centre using space-filling representations. Part b is modified, with permission, from Ref. 191 © (2011) Genetics Society of America. Jaunās pieejas augu genoma modifikācijai TALE faktori Transcription activator like effectors (TALEs) (Boch et al. 2009; Moscou and Bogdanove 2009). TALEs are produced by plant pathogens in the genus Xanthomonas, which deliver the proteins to plant cells during infection Proteīna-DNS mijiedarbības specifiskums TALE efektorā Christian M. et al., Targeting DNA Double-Strand Breaks with TAL Effector Nucleases, Genetics 186: 757–761 (October 2010) A schematic representation of a transcription activator-like effector nuclease (TALEN) pair. a | Each TALEN is composed of transcription activator-like effectors (TALEs) at the amino terminus and the FokI nuclease domain at the carboxyl terminus. Each TALE repeat is comprised of 33–35 amino acids and recognizes a single base pair through the amino acids at positions 12 and 13, which is called the repeat variable diresidue (RVD; shown in red). Target sequences of TALEN pairs are typically 30–40 bp in length, excluding spacers. b | In the TALE–DNA co-crystal structure, the RVDs in TALE interact with DNA in the major groove. The amino-terminal repeats (designated as 0 and −1 in the box) contact 5′ thymine. Part b is modified, with permission, from Ref. 73 © (2012) American Association for the Advancement of Science. Jaunās pieejas augu genoma modifikācijai CRISP-Cas CRISPR (clustered regularly interspaced short palindromic repeat) Cas (CRISPR-associated) genes, CRISPR-based adaptive immune systems Terns and Terns, 2011 Jaunās pieejas genoma modifikācijai CRISP-Cas Mali P. et al. RNA-Guided Human Genome Engineering via Cas9. Science, V339, p. 824, 2013 Schematic representations of RNA-guided engineered nucleases (RGENs). a | An RGEN is comprised of CRISPR (clustered regularly interspaced short palindromic repeat)-associated protein 9 (Cas9), a CRISPR RNA (crRNA) and a transactivating crRNA (tracrRNA), which form the dualRNA–Cas9. b | Alternatively, an RGEN can contain Cas9 and a single-chain guide RNA (sgRNA). The guide sequence in the crRNA (part a) or sgRNA (part b) is complementary to a 20-bp target DNA sequence known as protospacer, which is next to the 5′-NGG-3′ (where N represents any nucleotide) sequence known as protospacer adjacent motif (PAM). Grey dots indicate weak bonding. c | Target DNA cleaved by an RGEN yielding blunt ends is shown. d | A three-dimensional model of Cas9 complexed with DNA is shown. Part d courtesy of D. W. Taylor (University of California, Berkeley, USA), J. A. Doudna (University of California, Berkeley, USA) and M. Jinek (University of Zurich, Switzerland). J. J. Day (2014) New approaches to manipulating the epigenome. www.dialogues-cns.org DNS sekvenēšanas metožu attīstība DNS sekvenēšana ar daļēji specifiskas ķīmiskās degradācijas palīdzību Walter Gilbert, 1932 Andrejs Mirzabekovs, 1937 -2002 http://nationaldiagnostics.com/article_info.php/articles_ id/20 Frederick Sanger, 1918 www.nwfsc.noaa.gov/.../figur es/moranfig4.htm METODES Manuāla sekvenēšana Analīzes metodes Klasiskā pieeja: fragmentu klonēšana, subklonēšana, sekvenēšana http://seqcore.brcf.med.umich.edu/doc/ JAC, BAC, PAC, PUC Bioinformātikas idejas: nejauša klonēšana (shotgun), sekvenēšana http://img4.wikia.nocookie.net K. Venters, B.Klintons, F. Kolins, 2000, cilvēka genoma projekta finiša taisnē http://www.vfa-bio.de/static/ Resekvenēšanas metodes “Resekvenēšana”, vai genoma sekvenēšana nto reizi, vai genoma daļas sekvenēšana organismam, kam viena genoma sekvence jau zināma (vai pat radniecīgam organismam) ir vieglāka un lētāka nekā de novo sekvenēšana. Vairākas firmas piedāvā liela apjoma, ātrdarbīgas paralēlās resekvenēšanas platformas. 454 Life Sciences (http://www.454.com/enabling-technology/the-system.asp) Solexa (Illumina) (http://www.illumina.com/pages.ilmn?ID=203) PERSONISKIE GENOMA PROJEKTI Samuel Levy, et al. (Craig Venter) The Diploid Genome Sequence of an Individual Human PLoS BIOLOGY October 2007 | Volume 5 | Issue 2113 10 | e254 David A. Wheeler, et al. (James Watson) The complete genome of an individual by massively parallel DNA sequencing Nature 452, 872-876 (17 April 2008) Jeffrey M. Kidd, et al. Mapping and sequencing of structural variation from eight human genomes Nature 453, 56-64 (1 May 2008) Analīzes metodes Analīzes metodes Solex – Illumina tehnoloģija Analīzes metodes Solex – Illumina tehnoloģija Analīzes metodes Solex – Illumina tehnoloģija Solex – Illumina JONU PUSVADĪTĀJU SEKVENĒŠANA –ION TORRENT TECHNOLOGIES January 10, 2012 Life Technologies Benchtop Ion Proton Sequencer will sequence human genomes in one day for less than $1000 by yearend and Illumina will have a competing sub-$1000 per human genome sequencer by yearend The Ion Proton™ Sequencer is ideal for sequencing both exomes — regions in the DNA that code for protein — and human genomes. The Ion Proton™ I Chip, ideal for sequencing exomes, will be available mid-2012. The Ion Proton™ II Chip, ideal for sequencing whole human genomes, will be available about six months later. In addition, the Ion Proton™ OneTouch™ system automates template prep and a standalone Ion Proton™ Torrent Server performs the primary and secondary data analysis. Nanopore DNA sequencing technique promises entire genome in minutes or your money back https://www.sciencenews.org/sites/ http://www.futuretimeline.net/blog/images/1137.jpg Rick Merritt DNA Chip Will Plug Into Handsets 6/9/2014 Sekvenēšanas perspektīvas