The next generation of CRISPR–Cas technologies and applications

A Pickar-Oliver, CA Gersbach - Nature reviews Molecular cell biology, 2019 - nature.com
The prokaryote-derived CRISPR–Cas genome editing systems have transformed our ability
to manipulate, detect, image and annotate specific DNA and RNA sequences in living cells …

Editing the genome without double-stranded DNA breaks

AC Komor, AH Badran, DR Liu - ACS chemical biology, 2018 - ACS Publications
Genome editing methods have commonly relied on the initial introduction of double-
stranded DNA breaks (DSBs), resulting in stochastic insertions, deletions, and translocations …

Programmable base editing of A• T to G• C in genomic DNA without DNA cleavage

NM Gaudelli, AC Komor, HA Rees, MS Packer… - Nature, 2017 - nature.com
The spontaneous deamination of cytosine is a major source of transitions from C• G to T• A
base pairs, which account for half of known pathogenic point mutations in humans. The …

A large panel of isogenic APP and PSEN1 mutant human iPSC neurons reveals shared endosomal abnormalities mediated by APP β-CTFs, not Aβ

D Kwart, A Gregg, C Scheckel, EA Murphy, D Paquet… - Neuron, 2019 - cell.com
Familial Alzheimer's disease (fAD) results from mutations in the amyloid precursor protein
(APP) and presenilin (PSEN1 and PSEN2) genes. Here we leveraged recent advances in …

In vivo base editing of post-mitotic sensory cells

WH Yeh, H Chiang, HA Rees, ASB Edge… - Nature …, 2018 - nature.com
Programmable nucleases can introduce precise changes to genomic DNA through
homology-directed repair (HDR). Unfortunately, HDR is largely restricted to mitotic cells, and …

Loss of TREM2 rescues hyperactivation of microglia, but not lysosomal deficits and neurotoxicity in models of progranulin deficiency

A Reifschneider, S Robinson, B van Lengerich… - The EMBO …, 2022 - embopress.org
Haploinsufficiency of the progranulin (PGRN)‐encoding gene (GRN) causes frontotemporal
lobar degeneration (GRN‐FTLD) and results in microglial hyperactivation, TREM2 …

TPC2 rescues lysosomal storage in mucolipidosis type IV, Niemann–Pick type C1, and Batten disease

A Scotto Rosato, EK Krogsaeter, D Jaślan… - EMBO molecular …, 2022 - embopress.org
Lysosomes are cell organelles that degrade macromolecules to recycle their components. If
lysosomal degradative function is impaired, eg, due to mutations in lysosomal enzymes or …

Homozygous might be hemizygous: CRISPR/Cas9 editing in iPSCs results in detrimental on-target defects that escape standard quality controls

D Simkin, V Papakis, BI Bustos, CM Ambrosi, SJ Ryan… - Stem Cell Reports, 2022 - cell.com
The ability to precisely edit the genome of human induced pluripotent stem cell (iPSC) lines
using CRISPR/Cas9 has enabled the development of cellular models that can address …

Detection of deleterious on-target effects after HDR-mediated CRISPR editing

I Weisheit, JA Kroeger, R Malik, J Klimmt, D Crusius… - Cell reports, 2020 - cell.com
CRISPR genome editing is a promising tool for translational research but can cause
undesired editing outcomes, both on target at the edited locus and off target at other …

Development of hRad51–Cas9 nickase fusions that mediate HDR without double-stranded breaks

HA Rees, WH Yeh, DR Liu - Nature communications, 2019 - nature.com
In mammalian cells, double-stranded DNA breaks (DSBs) are preferentially repaired
through end-joining processes that generally lead to mixtures of insertions and deletions …