2017 CHOC Research Week – Call for Abstracts

Please submit your abstract for CHOC Research Week to jhuang@choc.org by Friday, October 27th, 5PM PT.



  1. Each presenter is limited to one abstract submission.
  2. 2.  Identify the Presenting Author, Institute & Department.
  3. Abstracts are limited to 300 words. Abstracts that have previously been presented at other meetings may be submitted for Research Week.
  4. State the Primary Topic and Secondary Topic of your abstract and provide 1-5 Keywords to describe your abstract.

Questions? Contact Jeff Huang at (714) 509-7915 [x17915] or jhuang@choc.org


Presenting Author:  Jeffrey Y Huang, PhD
Institute: CHOC Children’s Research Institute (Orange, CA)
Department: Metabolic Research
Primary Topic:  Lysosomal storage disorder
Secondary Topic: CRISPR-Cas9 genome editing
Keywords:  Pompe disease, lysosome, glycogen, animal model

Engineering Pompe disease models using CRISPR-Cas9 genome editing

Jeffrey Y Huang, PhD1; Abhinav Grover, MD1,2; and Raymond Y Wang, MD1

1CHOC Children’s Research Institute, Orange, CA 92868
2Chapman University School of Pharmacy, Irvine, CA 92618


Introduction: Pompe disease (PD) is a serious and progressive disorder caused by a genetic deficiency in acid alpha glucosidase (GAA) – an enzyme that recycles stored muscle cell glycogen for energy.  PD patients with near-complete GAA deficiency – infantile-onset PD - present within the first few months of life with severe heart enlargement, heart failure, weakness of the muscles, respiratory failure, and demonstrate rapidly progressive, fatal disease.  Without treatment, the average age that infantile-onset PD patients require permanent breathing support is about 6 months; the average age of death is about 9 months.


Project Aim: There are currently no models of PD featuring single nucleotide GAA mutations – which may be amenable to personalized genome editing.  The goal of this project is to engineer PD-specific GAA mutations in C2C12 cells - a mouse myoblast line.  We will seek to introduce single nucleotide GAA mutations homologous to common mutations found in infanitile-onset PD using CRISPR-Cas9 genome editing. This set of experiments will allow us to determine the efficacy and specificity of our CRISPR-Cas9 system in a cultured cell line prior to in vivo testing.


Results/Conclusion: Our GAA-specific CRISPR-Cas9 system led to the following results in C2C12 cells: a) GAA c.1826insA gRNA [Infantile-onset PD CRIM-] introduced the expected 1bp insertion knock-in mutation, b) GAA c.1935C>A gRNA [Infantile-onset PD CRIM+] introduced the expected C>A knock-in mutation. Further analysis showed that there is no detectable Cas9 activity at the 10 off-target sites most similar to the GAA c.1826 gRNA target sequence. Future experiments will aim to examine GAA c.1935 gRNA off-target sites and design and test repair gRNAs/templates for genomic correction of knock-in cell lines.