Advances in Gene Editing

Targeted genome editing in human repopulating haematopoietic stem cells

• Pietro Genovese,
• Giulia Schiroli,
• Giulia Escobar,
• Tiziano Di Tomaso,
• Claudia Firrito,
• Andrea Calabria,
• Davide Moi,
• Roberta Mazzieri,
• Chiara Bonini,
• Michael C. Holmes,
• Philip D. Gregory,
• Mirjam van der Burg,
• Bernhard Gentner,
• Eugenio Montini,
• Angelo Lombardo
• & Luigi Naldini

• Affiliations
• Contributions
• Corresponding author

Nature

 

(2014)

 

doi:10.1038/nature13420

Received

 

02 August 2013 

Accepted

 

29 April 2014 

Published online

 

28 May 2014

Abstract

• Abstract• 
• References• 
• Author information• 
• Extended data figures and tables• 
• Supplementary information

Targeted genome editing by artificial nucleases has brought the goal of site-specific transgene integration and gene correction within the reach of gene therapy. However, its application to long-term repopulating haematopoietic stem cells (HSCs) has remained elusive. Here we show that poor permissiveness to gene transfer and limited proficiency of the homology-directed DNA repair pathway constrain gene targeting in human HSCs. By tailoring delivery platforms and culture conditions we overcame these barriers and provide stringent evidence of targeted integration in human HSCs by long-term multilineage repopulation of transplanted mice. We demonstrate the therapeutic potential of our strategy by targeting a corrective complementary DNA into the IL2RG gene of HSCs from healthy donors and a subject with X-linked severe combined immunodeficiency (SCID-X1). Gene-edited HSCs sustained normal haematopoiesis and gave rise to functional lymphoid cells that possess a selective growth advantage over those carrying disruptive IL2RG mutations. These results open up new avenues for treating SCID-X1 and other diseases.

Field Trip: Gone Fishin'

Bertrand's class went on a field trip to Camp Kostopulos today.  Camp Kostopulos is a residential summer camp where kids, teens, and adults with disabilities are able to choose from two options--a five day residential camp or travel trips.  They also offer year-round recreational programs--such as horseback riding and fishing.

Bertrand (and Victoria) were able to fish with the help of student volunteers from the Open Classroom, a local charter school.  The students, teachers, and parents from Open Classroom were wonderful.  I think everyone from both schools had a great time.   :)

Everyone called him "Indiana" since he was dressed like Indiana Jones.  

Victoria got to hold and release a fish.  Highlight of her week.

This is how real fishing is done!  Zzzz...

Utah Renaissance Festival

This morning we went to the Utah Renaissance Festival & Fantasy Faire courtesy of Hope Kids.

Bertrand laughed every time he heard a horse whinny, and Victoria enjoyed riding a one!

We partook in the obligatory turkey leg

...followed up by tornado dogs, fried oreos, and italian ice.

(Bertrand is a fan of fried oreos.  Who knew?!)

We had to leave in a rush because Victoria spied Captain Hook.  

She was adamant that we "run"!

A pre-Captain Hook sighting video for the grandparents: Victoria riding the pony.

Biggest Brother Bertrand, Big Sister Victoria...

...and (soon) Baby Brother Winston

Ultrasound image, February 2014

With the due date of June 18, it's hard to believe that in less than a month he'll be here.

In some sense, Winston's gestation was much longer than the typical 9 months--his "conception" began almost 24 months ago in June 2012.

He is a baby that science made possible.

The process involved acronyms like IVF, PGD, CVS and many laboratories: Duke University, University of Utah, Natera, Counsyl, and GeneDx.

(And, needles.  Lots of needles.)

It was a difficult process, physically and emotionally.  There was a lot of anxiety, failure, and loss.

It was worth it.

We know that he isn't affected by N-glycanase deficiency--in fact, he's not even a carrier.

He is also not a carrier for 107 other rare genetic conditions.

And, he is a 100% chromosome normal XY male.

We got lucky.

_____________________________________________________
This morning, Bertrand and Victoria attended their "Sibling Preparation Class" at the hospital where mommy will deliver.  The class involved anatomy posters, sibling books, baby dolls, coloring sheets, a brief video, and a hospital tour.  Bertrand, being the old hand that he is, was a bit bored.  Victoria was riveted and kept interrupting to lecture the teacher (a L&D nurse) on babies... and Jake and the Neverland Pirates.  So, it went well.  :)  We are all ready and excited to meet baby Winston.

With Daddy in the hospital lobby.

Looking for the Nursery.

Bertrand's expression says, "I know this already!"

The attentive pupil.

These two take baby visitation very seriously.

2014 Epilepsy Education Conference

Get Seizure Smart! Educational ConferenceJoin us for the 7th Annual Get Seizure Smart! Educational Conference.

June 7th, 2014
8am to 4pm
Little America
500 S Main St
Salt Lake City, UT
84101 

To Register - please fill out this form and remit payment.

"Change is the end result of all true learning."
- Leo Buscaglia

This past year, the changes that have occurred in the epilepsy community include new diagnostic tools, new treatments and more research into what causes epilepsy and what can help stop seizures. The EAU is pleased to have some of the experts in this field of study and practice, speaking at our annual Get Seizure Smart! Education Conference.

Some of the respected speakers include:

Keynote Address:

---

Genetics of Epilepsy: The Role of Dravet Syndrome
Dr. Ian Miller - Miami Children's Hospital


Special Guest Speakers:

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Cannabidiol for Seizure Management
Heather Jackson - Executive Director - Realm of Caring

Epilepsy 101 for Adults
Dr. Jeffrey Bigelow, MD, MPH - Neurologist at Intermountain Medical Center

Epilepsy 101 for Pediatrics
Dr. Matthew Sweney, Assistant Professor, Pediatrics and Neurology - Primary Children's Hospital

The State of Cannabis Oil
Jennifer May, Co-Founder: Hope 4 Children with Epilepsy,

The genetics of epilepsy and personalized medicine: What does the future look like?
Dr. Reid Robison, Chief Executive Officer, Tute Genomics Inc.


Honored Speakers:

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The Practical Application of Chiropractics for Epilepsy
Dr. K.C. Oliver - Board Certified Chiropractic Neurologist, Founding Partner - The Neuro Clinic in Lehi UT

Trusts and Living Wills
Jared Allebest - Adjunct Professor, UVU - Owner, Allebest Law Group

Empowering Parents: Caregiver resources and how to use them.
Kim Orton - Pediatric Epilepsy Care Coordinator for the University of Utah, School of Medicine at Primary Children's Hospital.

Funding, Medicaid and SSI
Jodi Hansen - Utah Parent Center and Utah Family Voices.

Neuropsychological, Behavioral, and Social Challenges Associated with Epilepsy
Dr. Jenise Jensen - Pediatric Neuropsychologist

My Personal Experience and Accepting Epilepsy
Chris Connor - Youth Leader, Epilepsy Association Board of Directors

Essential Oils and Mental Health
Leslie Reader - Wellness Advocate 

Upsee: First Impressions

Bertrand's expression here: "Whoa! I'm standing up!"

The Upsee is a "mobility device that lets children with motor impairment stand and walk with the help of an adult".  It is a brand new product that is all the rage in the special needs community--and for good reason!

We've had the Upsee for over a week, and I can't say enough good things about the product or the company.

As I see it, Firefly by Leckey, the maker of the Upsee, is the Apple of special needs products.  The marketing, branding, and packaging of the Upsee have all been top notch.  (The unboxing of the Upsee was glorious!)  More importantly, the product itself is of the highest quality, intuitive to use, and supremely effective.

At $489, the Upsee is by no means an inexpensive product but, compared to other special needs devices, it is a bargain!  (A wheelchair like Bertrand's will run $8,000, and other gait devices run upwards of $3,000.)

Bertrand has really enjoyed being in the Upsee, and is building up his endurance.  We plan on sending it to school with him daily so that his classmates may also benefit from using it.

Field Trip: Little Dell Reservoir

Little Dell Reservoir

Today, Bertrand's class had a field trip to Little Dell Reservoir to go canoeing courtesy of Splore, a great local organization that provides opportunities for adventure to people with physical and/or cognitive disabilities.  And, an adventure it was!  The entire class--including Victoria--had a fantastic time.  :)

On the school bus and ready to go!

Headed down to the boat ramp.

The very first canoe to be loaded and pushing off.

Victoria serves as scout, pointing the way.

Canoeing on a very windy day.

Paddling further out.

Pointing at Mama back on shore.

Paddling out to see some Canada Geese.

Being joined by a classmate in another canoe.

On the way back to shore.

Bertrand was singing a happy song.

Docking and unloading the canoe.

A very happy boater.

Canoeing wiped us out!

Can I go again?!

Surrounded by classmates/friends.

Little sister photobomb.

Boy, was canoeing fun or what?!

Can't wait to do this again next year!

Animals Inside Out

Today, Bertrand left school a bit early to see the "Animals Inside Out" exhibit at The Leonardo, a local science museum.

Bertrand could see every exhibit unimpeded from his wheelchair since we went at an off-peak time.

Both kids loved the exhibit!

Victoria's most recent fascination has been with skeletons, muscles, and fossils.  Now, thanks to this exhibit, we can add organs (heart, lungs, stomach, etc.) to that list.

It's hard to see from these photos, but this bull is huge.  Bertrand was impressed.

The future doctor (PhD? MD? both?) examines her patient/research subject.

This giraffe was a favorite.

After the exhibit, the kids wanted a snack, so we hit the patisserie across the street.

Victoria cheerfully shared her "eclair fraiser" with Buddy.  

(He also polished off a third of mommy's kouign-amann bread pudding.)

As we left, Victoria declared the outing "a great adventure!"  We all agreed.  :)