HD journal presentation Godinho et al, Delivering a disease- - - PowerPoint PPT Presentation

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Dec 03, 2023 410 likes •556 views

Background for HD journal presentation Godinho et al, Delivering a disease- modifying treatment for Huntingtons disease, Drug Discovery Today , 2014 Abstract The human HTT gene and protein RNA interference (RNAi): short hairpin (SH) RNAs or

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Background for HD journal presentation

Godinho et al, Delivering a disease- modifying treatment for Huntington’s disease, Drug Discovery Today, 2014

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Abstract

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The human HTT gene and protein

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RNA interference (RNAi): short hairpin (SH) RNAs or short interfering (si)RNAs

Pol II = RNA polymerase II; pri-miRNA = primary miRNA; Note: some pri-miRNAs generated by pol III; Pasha = DGCR8 (DiGeorge syndrome critical region 8); Drosha = RNase III activity-containing protein ; Pre-miRNA = precursor miRNA; Dicer = RNAase III enzyme; RISC = RNA-induced silencing complex; RITS = RNA induced transcriptional silencing

“microRNA” Pathway “mirtron” Pathway

“Micro- processor Complex” RNA editing (not shown)

Pol II Pol II

“Guide” strand “Passenger” strand

RISC complex

argonaute (Ago)

shRNAs synthetic siRNAs

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Antisense oligonucleotides (ASOs)

PNA = peptide nucleic acid Morpholino: morpholine ring replaces ribose ring; phosphorodiamidate groups replace phosphate groups

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Ribozymes

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DNA enzymes

Schlosser K and Li Yingfu, Biologically inspired synthetic enzymes made from DNA, Chemistry and Biology 16, 311-322, 2009

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Genome editing: repair of double-stranded DNA breaks

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Genome editing: Zinc finger nucleases (ZFNs)

Carroll D, Genome Engineering with Zinc-finger nucleases, Genetics 188, 2011

Fok I restriction endonuclease

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Genome editing: Transcription activator-like nucleases (TALENs)

Each 34-amino acid repeat recognizes

ne base pair in double-stranded DNA

NTF3 = neurotrophin 3; CCR5 = chemokine C-C motif receptor 5 Boch J, 2011

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Genome editing: Clustered regulatory interspaces short palindromic repeat (CRISPR-Cas9) systems

CRISPR-Cas9 RCas9

Jennifer Doudna, Ph.D. Professor of Chemistry and Molecular and Cell Biology at the University of California, Berkeley Emmanuelle Charpentier, Ph.D. Department Head: Regulation in Infection Biology, Helmholtz Centre for Infection Research, Professor, Hannover Medical School

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Technical and ethical issues:

1. Off target effects
2. Efficient delivery to cells and tissues

(e.g. adeno-associated virus (AAV)-based; lipid-formulated siRNAs; nanoparticles)

3. Use for the modification of plants and animals
4. Use to treat or prevent human genetic diseases

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Recent internet headlines:

(Received $3,000,000 USD!)

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References

1. Godinho B, et al, Delivering a disease-modifying treatment for Huntington’s

disease, Drug Discovery Today 20, 50-64, 2015

2. Gupta RM and Musunuru K, Expanding the genetic editing tool kit: ZFNs,

TALENs, and CRISPER-Cas9, Journal Clinical Investigations 124, 4154-4161, 2014

3. Carroll D, Genome Engineering with zinc-finger nucleases, Genetics 188,

773-382, 2011

4. Miller JC, et al. A TALE nuclease architecture for efficient genome editing,

Nature Biotechnology 29, 143-148, 2011