Homology-directed gene repair Sequence correction xx exogenous repair substrate Rad51… sister chromatids I-SceI.

Slides:

Advertisements

Similar presentations

The Cell Cycle These notes are an introduction to The Cell Cycle Unit.

Advertisements

Gfp - Gene correction in trans: a model for gene therapy Luther Davis Jason Cummings Nancy Maizels.

How we can achieve disruption of the genes essential for cell viability in DT40 cells? Tackling essential genes; tagging Minoru Takata Laboratory of DNA.

Viruses and Gene Therapy

Zoo-352 Principles of genetics Lecture 3 The cell cycle and its checkpoints.

Andrew J. Pierce TOX 780 Single and Double-strand Break Repair TOX 780 Andrew Pierce Microbiology, Immunology and Molecular Genetics Toxicology University.

Cloning GADD45a into pCMV-Myc1 or pCMV-nV-His6 Gadd45EcoVF ATCCATGACTTTGGAGGAATTCTCGGCTGGAGAGC Gadd45XbaR GTAATCTAGATCACCGTTCAGGGAGATTAATCACTGG Cloning.

“CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system Thank you. Nucleeases will be used as a new tool that.

The analysis of homologous recombination frequency 1.There are two assays (artificial substrates) for double-strand break (DSB) induced homologous recombination.

DNA Organization, Replication, & Repair. Model for the structure of the nucleosome.

Instability: Mutation and DNA repair Mutations DNA repair.

Инвестиционный паспорт Муниципального образования «Целинский район»

(x – 8) (x + 8) = 0 x – 8 = 0 x + 8 = x = 8 x = (x + 5) (x + 2) = 0 x + 5 = 0 x + 2 = x = - 5 x = - 2.

Lentiviral Design for Delivery of Homing Endonucleases mCherry HA-NLS-HE RSVR U5 hPGK R U5sinU3 Repair Template Cppt RRE RRL Lentiviral Backbone.

Fundamentals of Biotechnology Lecture #07. Bacterial Artificial Chromosomes Many vectors which are popularly used for DNA cloning in bacterial cells contain.

Monnat Lab progress monomerizations of Cre, Mso (Hui) 2 nd round designs vs. Anopheles targets (Umut) in vivo recombination reporter assays (Ray, Hui)

The chicken light chain locus: a model of homology-directed repair of SSBs? VV VJ C VJ C Chicken pre-B cell line DT4O: carries out constitutive templated.

Section 11.3 Genetic Changes.

I-SceI-mediated Genome Editing in the Canine Model Kiem lab Applications Meeting Feb 23, 2009.

Describe what you see, why do you think this occurs and how?

NGEC Applications Meeting Mike Certo Scharenberg Lab.

Determine the sequence of genes along a chromosome based on the following recombination frequencies A-C 20% A-D 10% B-C 15% B-D 5%

Part 2. Cell reproduction of somatic cells (all cells except sperm/egg) This is how we grow, develop, and repair Involves chromosomes: Complex structure.

Homology-directed gene repair

May 2, 2011 Of the two causes we looked at Friday, which do you think is most likely responsible for frog malformations? Why? How many chromosomes do humans.

Restriction enzymes Are found in bacteria and are used to cut up DNA from a virus that might enter and take over the bacteria. They cut at specific sequences.

Homologous Recombination

照片档案整理一、照片档案的含义二、照片档案的归档范围三、卷内照片的分类、组卷、排序与编号四、填写照片档案说明五、照片档案编目及封面、备考填写六、数码照片整理方法七、照片档案的保管与保护.

공무원연금관리공단 광주지부 공무원대부등 공적연금 연계제도 공무원연금관리공단 광주지부. 공적연금 연계제도 국민연금과 직역연금 ( 공무원 / 사학 / 군인 / 별정우체국 ) 간의 연계가 이루어지지 않고 있 어 공적연금의 사각지대가 발생해 노후생활안정 달성 미흡 연계제도 시행전.

Жюль Верн ( ). Я мальчиком мечтал, читая Жюля Верна, Что тени вымысла плоть обретут для нас; Что поплывет судно громадней «Грейт Истерна»; Что.

Provider of Global Contract Research Services Accelerating Preclinical Research, Drug Discovery & Therapeutics Services > Generation of Stable Cell Lines.

Why Do Cells Divide?. Why Do Cells Divide? 1. Surface area to volume ratio Efficiency of moving materials into the cell Efficiency of moving materials.

Cell Division.  Grow  DNA Replication  Prepares to divide  Prep stage (90% of the cell life.)

CHAPTER 11-4 VOCAB.

Terms YOU need to know!.

Instability: Mutation and DNA repair

Why Do Cells Divide?.

Limits to Cell Growth There are two main reasons why cells divide rather than continuing to grow indefinitely. The larger a cell becomes, the.

NEW UNIT! GENETICS.

BIOLOGY Unit 4 Notes: Mitosis

Figure 2 Use of CRISPR/Cas9 for genome editing

What does the word Promoter mean?

Supplemental Figure S1 a b

Volume 55, Issue 1, Pages (July 2014)

Unit 3 Chapter 10 Cell Cycle

Some of the “C” words.

Expression of SYCE2 activates the DSB repair pathway.

Cell Division: Meiosis

Volume 17, Issue 7, Pages (April 2007)

MCM9 Is Required for Mammalian DNA Mismatch Repair

Volume 1, Issue 3, Pages (September 2013)

Volume 14, Issue 6, Pages (February 2016)

Instability: Mutation and DNA repair

BRCA2 Is Required for Homology-Directed Repair of Chromosomal Breaks

Molecular Roadblocks for Cellular Reprogramming

Somatic Hypermutation of Immunoglobulin Genes

Volume 24, Issue 3, Pages (March 2016)

Volume 28, Issue 3, Pages (November 2007)

Control of Sister Chromatid Recombination by Histone H2AX

True/False: Cell Size Limitations

Cell size is limited. Volume increases faster than surface area.

Efficient Sleeping Beauty DNA Transposition From DNA Minicircles

XRCC3 Controls the Fidelity of Homologous Recombination

Gene Amplification: Yeast Takes a Turn

MMEJ plays a dominant role in double-strand DNA break repair in L

Nuclease Target Site Selection for Maximizing On-target Activity and Minimizing Off- target Effects in Genome Editing Ciaran M Lee, Thomas J Cradick,

New Tools for Genome Modification in Human Embryonic Stem Cells

CCR5 enhances both HDR and SSA DNA repair.

Presentation transcript:

Homology-directed gene repair Sequence correction xx exogenous repair substrate Rad51… sister chromatids I-SceI

Homology-directed gene repair Sequence correction xx exogenous repair substrate Rad51… sister chromatids I-SceI

DRGFP reporter puro R I-SceISTOPBcgI puro R BcgI GFP + gene conversion CACA CACA

293T stable DRGFP (S8) no DNA pCMV-I-SceI 48 hrs 72 hrs

Direct Repeat Reporters: recombination in cis

293T DRGFP (cis) I-SceI DRGFP+I-SceI DRGFP

293T DRGFP-lacO (cis) DRGFPlacO DRGFPlacO+I-SceI

Exogenous Donor Reporter: recombination in trans

293T ZFGFP + iGFP (trans) ZFGFP+iGFPZFGFP+iGFP+I-SceI

293T ZFGFP-lacO + iGFP (trans) ZFGFP-lacOZFGFP-lacO+iGFP+I-SceI

Too Much DNA?

293T: Efficiency of Transient Transfection ng eGFP no DNA 1.0  g eGFP2.0  g eGFP4.0  g eGFP 62 ng eGFP125 ng eGFP250 ng eGFP

“Stable” DRGFP?

“Stable” DRGFP (puro S ) No DNA CMV-Sce 48 hrs73 hrs98 hrs

“Stable” DRGFP (puro R ) No DNA CMV-Sce 48 hrs73 hrs98 hrs

Plans Lines with integrated trans reporter –Enhance repair by tethering chromatin modifiers and repair factors Back to B-cells