advances in algal synthetic biology technologies synbi… · geneart® seamless /seamless plus...

1 | Life Technologies™ Proprietary and Confidential 12/19/2013

Advances in Algal Synthetic Biology Technologies

Farzad Haerizadeh

Pacific Rim Summit

December 10, 2013


Life Technologies at a glance

Revenue ~$3.8B

Countries 160

Employees ~ 10,000+

Patents, licenses 5,000+

Products ~50,000+

Citations 675,000+


Our Business

• Consumables and instruments used to sequence and analyze DNA and Gene Function

• Molecular Diagnostic Technologies for cancer, inherited genetic disease, and infectious disease

• Consumables and instrumentation for all types of sample preparation, molecular, and cellular research

• Synthetic Biology

• Cell culture media for basic research and drug development

• Bioproduction

• Forensic Testing

• Food Safety Testing

• Animal Health

Genetic Analysis and Medical Sciences

Applied Markets

Biosciences (Molecular Biology and Cell Analysis)

https://www.invitrogen.com/site/us/en/home/brands/Product-Brand/acrometrix.html


Synthetic Biology at LIFE

Various hosts


SynBio / Cell Engineering/ Complete Workflow

Understand/ Design

Build Payload/ Tools

Engineer Cells

Analyze Results

• Payload/ Delivery

• Chromosomal integration

• Genome Editing

• Genomic circuit design

• Gene regulation and

chromatin remodeling

• Understanding the

challenge

• Designing the solution(s)

• Refining predictions

• Synthesize DNA de novo

• Assembly from parts list

• Evolving new proteins

• Collating data to allow input

into metabolic models

• ‘Omic’ analysis of pre- and

post-engineered cells

Use modified cells


GeneArt® StringsTM DNA Fragments

• Custom-made linear, double-stranded

DNA fragments up to 1,000 bp in length,

ready for cloning in your lab.

• We offer three categories of Strings™

fragments:

• 100–600 bp

• 601–750 bp

• 751–1,000 bp

• 5–7 business days delivery


Gene Synthesis & Assembly Tools

1. GeneArt® High-Order Assembly System

• Yeast based, up to 110 kb constructs

• Oligo stitching (no end-homology required)

• Gene synthesis.

2. GeneArt® Seamless /Seamless Plus

Cloning/Assembly

• In vitro, up to 14 kb inserts, vector construction

• In vitro, up to 40 kb inserts/larger constructs

• Horizontal DNA transfer (no ex vivo DNA manipulation)

3. GeneArt® Site-Directed Mutagenesis Kit

• Up to 14 kb circular targets

• Single or multi-site editing

• Degenerated mutations allowed

4. DIY Gene Synthesis – CorrectASE™ Enzyme

• 4-day high fidelity gene synthesis workflow

• Significantly reduces oligonucleotide errors

6. GeneArt® Primer and Construct Design Tools

• Assembly and mutagenesis design tool

• GenBank annotated sequences

• Oligo export and ordering features

http://www.technoplas.com.au/images/StandardImage/4010_c.gif


GeneArt® Type IIs Assembly Kits

- For Difficult or Repetitive Sequences!

With GeneArt® Type IIs kits you can:

• Assemble single or multiple (up to 8) DNA fragments in any order, into any

compatible vector, without scars

• Avoid homologous recombination and associated rearrangements when cloning

homologous or repetitive sequences

• Perform your restriction/ligation reaction using any one of three Type IIs

enzymes (AarI, BsaI, and BbsI)

• Supported by GeneArt® Primer and Construct Design Tool for efficient assembly


Random Mutagenesis & Integration : no control on where the edits are made

RNAi Mediated Gene Knockdown:

•Post transcriptional hence a temporary effect on gene function

Targeted Integration via Homologous Recombination:

•Low efficiency and laborious downstream screening methods

First Generation Genome Editing tool

•Zinc Finger Technology: one ZnF repeat per 3 bases . Requires screening and selection .

Next Generation Genome Editing Tools

•TAL Effector Technology

• CRISPR/Cas Technology

Gain Precision and control

No control, No precision

Programmable DNA-binding proteins/ editing tools • Engineered to edit specific DNA sequence • Allow targeted gene manipulation

Efficiency, precision & control in cell engineering


GeneArt® Precision TALs

• Activation of transcription (vp16 or vp64)

– Increasing expression level of endogenous genes

• Steric repression & custom design (MCS)

– Target any locus in the genome with the effectors domain of your

choice

• Nuclease function (Fok1 Nuclease Pair)

– Silencing/Stable Knock-Outs

– Incorporation of exogenous DNA/Knock-Ins

• Repression of transcription (KRAB)

– Targeted repression of gene expression

GeneArt® Precision TALs provide custom DNA-binding proteins for

accurate DNA targeting and precise genome editing.


TAL effectors can be used to regulate gene expression

TAL-Repressor

Light-On Light-Off

• The indicated plasmids transfected into 293FT cells.

Images were taken 48h post-transfection.

pcDNA3

pCMV-mini GFP 3XTAL

TAL-Activator

pCMV-mini GFP

pcDNA3

PCMV GFP 2xTAL

PCMV GFP 2xTAL

X


• RNA-guided genome editing tool that is simple, easy and quick to implement.

• Allows you to edit and engineer the target of your choice from a single plasmid.

• GeneArt® CRISPR Nuclease reporter vector system expresses Cas9 as well as CRISPR RNA and

reporters from the same vector.

• Now available as a build-it-yourself ready to use kit or custom service option

Guide RNA (gRNA) which is a

chimera between target

complementary crRNA and

tracrRNA (constant component

not variable) forms a complex with

Cas9 nuclease.


Algal Technologies


Algae Research Tools and Capabilities


MAX Efficiency® Transformation

Reagent

•GeneArt® Synechococcus Protein

Expression Kit

•GeneArt® Chlamydomonas

Protein Expression Kit

GeneArt® Cryopreservation

Kit for Algae

Life’s Next Generation Algal Tools Just Launched!

Offering a complete solution


MAX Efficiency® Transformation

Reagent

•GeneArt® Synechococcus Protein

Expression Kit

•GeneArt® Chlamydomonas

Protein Expression Kit

GeneArt® Cryopreservation

Kit for Algae

Life’s Next Generation Algal Tools Just Launched!

• No need for liquid nitrogen

• No need for expensive

equipments

• No more genetic drifts

• No more contamination!

• And more………….

Chlamy

Chlorella

Common homemade

Reagent

800 fold


Going Forward

Develop synthetic biology tools and technologies

Comprehensive cell/ genome engineering workflow toolbox

Partnerships and services cell engineering

− Applying our toolbox to create novel solutions for basic research,

bioproduction, drug discovery, editing and … (?)

Design Payload Delivery Culture Characterize Measure


Thanks!

© 2013 Life Technologies Corporation. All rights reserved.

The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners.

• www.lifetechnologies.com/syntheticbiology

• [email protected]

• [email protected]

http://www.lifetechnologies.com/syntheticbiology

mailto:[email protected]



advances in algal synthetic biology technologies synbi… · geneart® seamless /seamless plus...

Documents