Source 1primary paper2018
Toolkit/LADL
LADL
Also known as: Light-activated dynamic looping, Light-activated dynamic looping system
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
LADL (light-activated dynamic looping system) is a multi-component optogenetic genome-engineering platform that targets two genomic anchors with CRISPR guide RNAs and promotes their spatial co-localization through light-induced heterodimerization between CRY2 and dCas9-CIBN. In the cited 2018 study, this engineered looping altered endogenous gene expression, including increased nascent Zfp462 transcription and increased synchronous Sox2 expression.
Usefulness & Problems
Why this is useful
LADL is useful for experimentally imposing specific chromatin contacts at endogenous loci with light as the control input. The reported application shows that induced looping can modulate transcriptional output and allelic synchrony of native genes.
Source:
Here we report a new class of 3-D optoepigenetic tools for the directed rearrangement of 3-D chromatin looping on short time scales using blue light.
Problem solved
LADL addresses the problem of how to causally test whether a defined genomic looping interaction is sufficient to alter endogenous gene expression. It provides a way to recruit two selected genomic anchors into proximity using CRISPR guide RNA targeting coupled to optogenetic protein association.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
Heterodimerizationlight-induced heterodimerizationlight-induced spatial co-localizationspatial co-localizationTechniques
No technique tags yet.
Target processes
editinglocalizationrecombinationInput: Light
Implementation Constraints
The reported design uses CRISPR guide RNAs to target two genomic anchors and relies on a dCas9-CIBN fusion together with CRY2 for light-responsive recruitment. The available evidence does not specify illumination parameters, delivery format, host cell system, or construct stoichiometry.
The supplied evidence comes from a single cited study and supports only the specific reported gene-expression outcomes and targeting mechanism. The evidence provided here does not report quantitative kinetics, reversibility, off-target effects, cell-type generality, or performance across many loci.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
LADL increases synchronous Sox2 expression after reinforcement of a known Sox2-SE looping interaction.
Moreover, LADL also increased synchronous Sox2 expression after reinforcement of a known Sox2 -SE looping interaction.
LADL increases synchronous Sox2 expression after reinforcement of a known Sox2-SE looping interaction.
Moreover, LADL also increased synchronous Sox2 expression after reinforcement of a known Sox2 -SE looping interaction.
LADL increases synchronous Sox2 expression after reinforcement of a known Sox2-SE looping interaction.
Moreover, LADL also increased synchronous Sox2 expression after reinforcement of a known Sox2 -SE looping interaction.
LADL increases synchronous Sox2 expression after reinforcement of a known Sox2-SE looping interaction.
Moreover, LADL also increased synchronous Sox2 expression after reinforcement of a known Sox2 -SE looping interaction.
LADL increases synchronous Sox2 expression after reinforcement of a known Sox2-SE looping interaction.
Moreover, LADL also increased synchronous Sox2 expression after reinforcement of a known Sox2 -SE looping interaction.
LADL increases synchronous Sox2 expression after reinforcement of a known Sox2-SE looping interaction.
Moreover, LADL also increased synchronous Sox2 expression after reinforcement of a known Sox2 -SE looping interaction.
LADL increases synchronous Sox2 expression after reinforcement of a known Sox2-SE looping interaction.
Moreover, LADL also increased synchronous Sox2 expression after reinforcement of a known Sox2 -SE looping interaction.
LADL increases total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
Using single molecule RNA FISH, we observe a LADL-induced increase in the total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
LADL increases total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
Using single molecule RNA FISH, we observe a LADL-induced increase in the total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
LADL increases total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
Using single molecule RNA FISH, we observe a LADL-induced increase in the total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
LADL increases total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
Using single molecule RNA FISH, we observe a LADL-induced increase in the total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
LADL increases total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
Using single molecule RNA FISH, we observe a LADL-induced increase in the total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
LADL increases total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
Using single molecule RNA FISH, we observe a LADL-induced increase in the total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
LADL increases total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
Using single molecule RNA FISH, we observe a LADL-induced increase in the total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
LADL targets two genomic anchors with CRISPR guide RNAs and engineers their spatial co-localization through light-induced heterodimerization of CRY2 with dCas9-CIBN.
We target our light-activated dynamic looping system (LADL) to two genomic anchors with CRISPR guide RNAs and engineer their spatial co-localization via light-induced heterodimerization of the cryptochrome 2 (CRY2) protein with dCas9-CIBN.
LADL targets two genomic anchors with CRISPR guide RNAs and engineers their spatial co-localization through light-induced heterodimerization of CRY2 with dCas9-CIBN.
We target our light-activated dynamic looping system (LADL) to two genomic anchors with CRISPR guide RNAs and engineer their spatial co-localization via light-induced heterodimerization of the cryptochrome 2 (CRY2) protein with dCas9-CIBN.
LADL targets two genomic anchors with CRISPR guide RNAs and engineers their spatial co-localization through light-induced heterodimerization of CRY2 with dCas9-CIBN.
We target our light-activated dynamic looping system (LADL) to two genomic anchors with CRISPR guide RNAs and engineer their spatial co-localization via light-induced heterodimerization of the cryptochrome 2 (CRY2) protein with dCas9-CIBN.
LADL targets two genomic anchors with CRISPR guide RNAs and engineers their spatial co-localization through light-induced heterodimerization of CRY2 with dCas9-CIBN.
We target our light-activated dynamic looping system (LADL) to two genomic anchors with CRISPR guide RNAs and engineer their spatial co-localization via light-induced heterodimerization of the cryptochrome 2 (CRY2) protein with dCas9-CIBN.
LADL targets two genomic anchors with CRISPR guide RNAs and engineers their spatial co-localization through light-induced heterodimerization of CRY2 with dCas9-CIBN.
We target our light-activated dynamic looping system (LADL) to two genomic anchors with CRISPR guide RNAs and engineer their spatial co-localization via light-induced heterodimerization of the cryptochrome 2 (CRY2) protein with dCas9-CIBN.
LADL targets two genomic anchors with CRISPR guide RNAs and engineers their spatial co-localization through light-induced heterodimerization of CRY2 with dCas9-CIBN.
We target our light-activated dynamic looping system (LADL) to two genomic anchors with CRISPR guide RNAs and engineer their spatial co-localization via light-induced heterodimerization of the cryptochrome 2 (CRY2) protein with dCas9-CIBN.
LADL targets two genomic anchors with CRISPR guide RNAs and engineers their spatial co-localization through light-induced heterodimerization of CRY2 with dCas9-CIBN.
We target our light-activated dynamic looping system (LADL) to two genomic anchors with CRISPR guide RNAs and engineer their spatial co-localization via light-induced heterodimerization of the cryptochrome 2 (CRY2) protein with dCas9-CIBN.
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors.
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors.
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors.
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors.
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors.
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors.
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors.
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors
LADL-induced looping changes occur as early as four hours after light induction.
Looping changes occur as early as four hours after light induction.
time to looping change 4 hours
LADL-induced looping changes occur as early as four hours after light induction.
Looping changes occur as early as four hours after light induction.
time to looping change 4 hours
LADL-induced looping changes occur as early as four hours after light induction.
Looping changes occur as early as four hours after light induction.
time to looping change 4 hours
LADL-induced looping changes occur as early as four hours after light induction.
Looping changes occur as early as four hours after light induction.
time to looping change 4 hours
LADL-induced looping changes occur as early as four hours after light induction.
Looping changes occur as early as four hours after light induction.
time to looping change 4 hours
LADL-induced looping changes occur as early as four hours after light induction.
Looping changes occur as early as four hours after light induction.
time to looping change 4 hours
LADL-induced looping changes occur as early as four hours after light induction.
Looping changes occur as early as four hours after light induction.
time to looping change 4 hours
LADL is a light-activated optoepigenetic tool for directed rearrangement of 3-D chromatin looping on short time scales using blue light.
Here we report a new class of 3-D optoepigenetic tools for the directed rearrangement of 3-D chromatin looping on short time scales using blue light.
LADL is a light-activated optoepigenetic tool for directed rearrangement of 3-D chromatin looping on short time scales using blue light.
Here we report a new class of 3-D optoepigenetic tools for the directed rearrangement of 3-D chromatin looping on short time scales using blue light.
LADL is a light-activated optoepigenetic tool for directed rearrangement of 3-D chromatin looping on short time scales using blue light.
Here we report a new class of 3-D optoepigenetic tools for the directed rearrangement of 3-D chromatin looping on short time scales using blue light.
LADL is a light-activated optoepigenetic tool for directed rearrangement of 3-D chromatin looping on short time scales using blue light.
Here we report a new class of 3-D optoepigenetic tools for the directed rearrangement of 3-D chromatin looping on short time scales using blue light.
LADL is a light-activated optoepigenetic tool for directed rearrangement of 3-D chromatin looping on short time scales using blue light.
Here we report a new class of 3-D optoepigenetic tools for the directed rearrangement of 3-D chromatin looping on short time scales using blue light.
LADL is a light-activated optoepigenetic tool for directed rearrangement of 3-D chromatin looping on short time scales using blue light.
Here we report a new class of 3-D optoepigenetic tools for the directed rearrangement of 3-D chromatin looping on short time scales using blue light.
LADL is a light-activated optoepigenetic tool for directed rearrangement of 3-D chromatin looping on short time scales using blue light.
Here we report a new class of 3-D optoepigenetic tools for the directed rearrangement of 3-D chromatin looping on short time scales using blue light.
Approval Evidence
1 source6 linked approval claimsfirst-pass slug ladl
We target our light-activated dynamic looping system (LADL) to two genomic anchors with CRISPR guide RNAs and engineer their spatial co-localization via light-induced heterodimerization of the cryptochrome 2 (CRY2) protein with dCas9-CIBN.
Source:
gene expression effectsupports
LADL increases synchronous Sox2 expression after reinforcement of a known Sox2-SE looping interaction.
Moreover, LADL also increased synchronous Sox2 expression after reinforcement of a known Sox2 -SE looping interaction.
Source:
gene expression effectsupports
LADL increases total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
Using single molecule RNA FISH, we observe a LADL-induced increase in the total nascent Zfp462 transcripts and the number of Zfp462 alleles expressing simultaneously per cell.
Source:
mechanismsupports
LADL targets two genomic anchors with CRISPR guide RNAs and engineers their spatial co-localization through light-induced heterodimerization of CRY2 with dCas9-CIBN.
We target our light-activated dynamic looping system (LADL) to two genomic anchors with CRISPR guide RNAs and engineer their spatial co-localization via light-induced heterodimerization of the cryptochrome 2 (CRY2) protein with dCas9-CIBN.
Source:
population level effectsupports
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors.
LADL facilitates loop synchronization across a large population of cells without exogenous chemical cofactors
Source:
timingsupports
LADL-induced looping changes occur as early as four hours after light induction.
Looping changes occur as early as four hours after light induction.
Source:
tool capabilitysupports
LADL is a light-activated optoepigenetic tool for directed rearrangement of 3-D chromatin looping on short time scales using blue light.
Here we report a new class of 3-D optoepigenetic tools for the directed rearrangement of 3-D chromatin looping on short time scales using blue light.
Source:
Comparisons
Source-backed strengths
The system acts at endogenous genomic sites specified by CRISPR guide RNAs and uses light-induced CRY2-dCas9-CIBN association to drive spatial co-localization. In the cited study, reinforcement of a Sox2-super-enhancer loop increased synchronous Sox2 expression, and a redirected interaction increased total nascent Zfp462 transcripts and the number of simultaneously expressing Zfp462 alleles per cell.
Ranked Citations
- 1.