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First-pass extracted concept

CRISPR/Cas9 AAVS1-targeted transgene integration

Candidate: toolkit itemType: engineering method1 source documents2 linked claims
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Aliases

AAVS1 safe harbor targeting, CRISPR/Cas9

Extracted Explainers

What the tool is doing

This approach inserts optogenetic or chemogenetic receptor genes into human iPSCs by CRISPR/Cas9 targeting of the AAVS1 safe-harbor locus. In this paper, integration succeeded in both HEK293T cells and iPSCs.

Source 1DOIPubMed

Resources required

The abstract supports a need for CRISPR/Cas9 editing targeted to AAVS1 and receptor transgene constructs such as ChR2 or hM4Di in human iPSCs. It also implies downstream methylation analysis and expression testing were used to evaluate outcomes.

Source 1DOIPubMed

What problem it solves

It addresses the need for efficient gene editing to construct modifiable iPSC-derived neurons carrying optogenetic or chemogenetic receptors.

Source 1DOIPubMed

What it does not solve

It does not ensure transgene expression after integration in human iPSCs, because the abstract reports silencing despite successful insertion.

Source 1DOIPubMed

Alternatives

The paper contrasts this approach with PiggyBac transposon-mediated integration.

Source 1DOIPubMed

Evidence Snippets

Here, we used CRISPR/Cas9 (targeting the AAVS1 safe harbor) ... to insert optogenetic and chemogenetic receptors (ChR2/hM4Di) into human iPSCs.
Evidence 1Source 1DOIPubMedprovenance

Supporting Sources

Linked Claims

Claim 1engineering outcomesupports2026Source 1DOIPubMed

CRISPR/Cas9 targeting AAVS1 and PiggyBac both enabled genomic integration of ChR2 and hM4Di transgenes in HEK293T cells and human iPSCs.

Quoted textsource-backed
While both systems successfully integrated genes into the genomes of HEK293T cells and iPSCs
Claim 2expression outcomesupports2026Source 1DOIPubMed

Despite successful genomic integration, receptor expression was detected only in HEK293T cells and not in human iPSCs.

Quoted textsource-backed
receptor expression was detected only in HEK293T cells