Review

grna cloning (Addgene inc)

Name: grna cloning
Brand: Addgene inc
Rating: 96 (441 reviews)

Addgene inc is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Addgene inc grna cloning
Grna Cloning, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 441 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/grna cloning/product/Addgene inc
Average 96 stars, based on 441 article reviews

grna cloning - by Bioz Stars, 2026-02

96/100 stars

Images

Image Search Results

CRISPR.BOT V1 setup steps. ( A ) Installation of the rail system for CRISPR.BOT V1. ( B ) The assembled version of the V1 robotic system using the LEGO digital designer program and the spare parts in the LEGO Mindstorms set. ( C ) Expansion of the rail system. ( D ) By expanding the bottom area of the rail system, a wider sub-floor is provided for the materials to be used in the designed experiments. ( E ) There are integrated colored papers for the plate carrier rail system and color sensor. When the color sensor encounters paper of the color specified in the coding, it activates the stop command to stop in the current position. ( F ) The position indicating that the syringe is aligned with the well center as determined by the coding.

Journal: Scientific Reports

Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications

doi: 10.1038/s41598-025-01655-2

Figure Lengend Snippet: CRISPR.BOT V1 setup steps. ( A ) Installation of the rail system for CRISPR.BOT V1. ( B ) The assembled version of the V1 robotic system using the LEGO digital designer program and the spare parts in the LEGO Mindstorms set. ( C ) Expansion of the rail system. ( D ) By expanding the bottom area of the rail system, a wider sub-floor is provided for the materials to be used in the designed experiments. ( E ) There are integrated colored papers for the plate carrier rail system and color sensor. When the color sensor encounters paper of the color specified in the coding, it activates the stop command to stop in the current position. ( F ) The position indicating that the syringe is aligned with the well center as determined by the coding.

Article Snippet: CRISPR + gRNA 1-2-3 Jurkat Cells (Clone E6-1, TIB-152, ATCC) in the 6-well plate were pulled at 100° and added to a whole 96-well plate. ( F ) Incubation of the 96-Well plate. ( G ) CRISPR.BOT-based subcloning of the transgenic cells. ( H ) Shows the positive and negative rates of GFP expression in cells sorted in the experiment performed with CRISPR.BOT.

Techniques: CRISPR

CRISPR.BOT V2 setup steps. ( A ) It is the mechanism that provides the movement of the syringe and is placed in the middle part of ( B ) The movement of the wheels is provided by toothed rail plates and the up-and-down movement of the straw is provided. ( C ) It is a part that keeps the syringe connected to the system. ( D ) Alternative parts are used to provide the syringe movement connected to the plunger. By placing the syringe plunger inside the red part, the movement of the part connected to the piston is provided by the movement of the impellers connected to the servo motor. With this system, the movement of the piston up and down is provided, and the fluid exchange process is carried out. ( E ) The prepared mechanism is placed in the pipette system. ( F ) Integration of the CRISPR.BOT pipetting system into the servo motor. The movement of the impeller connected to the large servo motor on the rail plate provides the movement of the pipette system, which performs the liquid exchange process. ( G ) Side view of the pipette system. The up-down movements of the pipette system are provided by the medium servo motor moving the wheels. ( H ) The gear piece enables the wheels to move on it. ( I ) CRISPR.BOT is the bottom part of the robotic system, called the frame. Image of the frame setup made from the LEGO digital designer program. ( J ) It is the bottom part of the robotic system, called the frame. The pipette system, which performs pipette movements and liquid exchange operations, is placed on the frame. Movements take place thanks to the toothed parts on the frame and the wheels at the bottom of the pipette system. The frame is fixed and does not have any mobility. ( K ) The system provides the liquid transition movements of the pipette between the other wells. The mechanism that provides the movement of the pipette system on the frame, was designed with the LEGO digital designer program. ( L ) It is the mechanism that enables the pipette system to move right, left, forward, and backward on the frame. The pipette provides fluid passage movements between other wells. ( M ) Pipette system that provides the right and left movements of the pipette.

Journal: Scientific Reports

Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications

doi: 10.1038/s41598-025-01655-2

Figure Lengend Snippet: CRISPR.BOT V2 setup steps. ( A ) It is the mechanism that provides the movement of the syringe and is placed in the middle part of ( B ) The movement of the wheels is provided by toothed rail plates and the up-and-down movement of the straw is provided. ( C ) It is a part that keeps the syringe connected to the system. ( D ) Alternative parts are used to provide the syringe movement connected to the plunger. By placing the syringe plunger inside the red part, the movement of the part connected to the piston is provided by the movement of the impellers connected to the servo motor. With this system, the movement of the piston up and down is provided, and the fluid exchange process is carried out. ( E ) The prepared mechanism is placed in the pipette system. ( F ) Integration of the CRISPR.BOT pipetting system into the servo motor. The movement of the impeller connected to the large servo motor on the rail plate provides the movement of the pipette system, which performs the liquid exchange process. ( G ) Side view of the pipette system. The up-down movements of the pipette system are provided by the medium servo motor moving the wheels. ( H ) The gear piece enables the wheels to move on it. ( I ) CRISPR.BOT is the bottom part of the robotic system, called the frame. Image of the frame setup made from the LEGO digital designer program. ( J ) It is the bottom part of the robotic system, called the frame. The pipette system, which performs pipette movements and liquid exchange operations, is placed on the frame. Movements take place thanks to the toothed parts on the frame and the wheels at the bottom of the pipette system. The frame is fixed and does not have any mobility. ( K ) The system provides the liquid transition movements of the pipette between the other wells. The mechanism that provides the movement of the pipette system on the frame, was designed with the LEGO digital designer program. ( L ) It is the mechanism that enables the pipette system to move right, left, forward, and backward on the frame. The pipette provides fluid passage movements between other wells. ( M ) Pipette system that provides the right and left movements of the pipette.

Techniques: CRISPR, Transferring

The schematization of bacterial transformation experiment steps and bacterial colonies after transformation using CRISPR.BOT. ( A ) Take 10–20 µl of DNA at an angle of 5° and add 50 µl of E.coli in the transwell at 4 °C. Then wait 30 min. ( B ) Take 60 µl of DNA + E.coli at an angle of 15° and wait for 20–30 s by immersing the pipette in 42 °C hot water. ( C ) Remove the pipette, which is immersed in 42 °C hot water, and leave E.coli + DNA in an 800 µl medium. Then shaking is done 3 times at intervals of 5 min for 15 min. ( D ) Take 205 µl 3 times at an angle of 40°, leave it in different parts of the Petri dish, and allow it to spread by shaking for 10 s. ( E ) Transgenic white bacterial colonies formed as a result of the transformation of plasmid DNA encoding GFP and Ampicillin (Amp) resistance gene into E.coli bacteria. µl, microliter; GFP, Green Fluorescent Protein; E.coli , Escherichia coli.

Journal: Scientific Reports

Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications

doi: 10.1038/s41598-025-01655-2

Figure Lengend Snippet: The schematization of bacterial transformation experiment steps and bacterial colonies after transformation using CRISPR.BOT. ( A ) Take 10–20 µl of DNA at an angle of 5° and add 50 µl of E.coli in the transwell at 4 °C. Then wait 30 min. ( B ) Take 60 µl of DNA + E.coli at an angle of 15° and wait for 20–30 s by immersing the pipette in 42 °C hot water. ( C ) Remove the pipette, which is immersed in 42 °C hot water, and leave E.coli + DNA in an 800 µl medium. Then shaking is done 3 times at intervals of 5 min for 15 min. ( D ) Take 205 µl 3 times at an angle of 40°, leave it in different parts of the Petri dish, and allow it to spread by shaking for 10 s. ( E ) Transgenic white bacterial colonies formed as a result of the transformation of plasmid DNA encoding GFP and Ampicillin (Amp) resistance gene into E.coli bacteria. µl, microliter; GFP, Green Fluorescent Protein; E.coli , Escherichia coli.

Techniques: Electroporation Bacterial Transformation, Transformation Assay, CRISPR, Transferring, Transgenic Assay, Plasmid Preparation, Bacteria

Genetic transfer in human cells, experiment programming and analysis of GFP protein expression. ( A ) Performing Cell Sowing, Jurkat Cells (Clone E6-1, TIB-152, ATCC) were cultivated in 4 wells at 2.5 × 10 4 cells/well. It was cultured in RPMI medium with a total volume of 500 µl. ( B ) Virus sample preparation, 1 ml of GFP (green fluorescent protein) Lentivirus was placed in one well of the 12-well plate. ( C ) Integration of Plates into the Autonomous Robot System, 12-well plates were placed in the frame of the Robot. ( D ) Addition of GFP Lentivirus to Cells, 70, 100, and 150° extractions were made from the well containing GFP Lentivirus (GFP LV) in a 12-well plate and added into wells 1, 2, and 3. ( E ) Incubation, after the robot performed the experiment, the 12-Well plate was placed in the incubator. ( F ) 70, 100, and 150° blocks were used for genetic transfer in human cells experiment programming. ( G ) 100°, and ( H ) 150° blocks that activate the pipette system are prepared in the experiment programming and perform the liquid withdrawal process. ( I ) Analysis of GFP protein expression in CRISPR.BOT modified transgenic Jurkat cell (Clone E6-1, TIB-152, ATCC) line using flow cytometry and ( J ) Fluorescence microscopy at 4X.

Journal: Scientific Reports

Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications

doi: 10.1038/s41598-025-01655-2

Figure Lengend Snippet: Genetic transfer in human cells, experiment programming and analysis of GFP protein expression. ( A ) Performing Cell Sowing, Jurkat Cells (Clone E6-1, TIB-152, ATCC) were cultivated in 4 wells at 2.5 × 10 4 cells/well. It was cultured in RPMI medium with a total volume of 500 µl. ( B ) Virus sample preparation, 1 ml of GFP (green fluorescent protein) Lentivirus was placed in one well of the 12-well plate. ( C ) Integration of Plates into the Autonomous Robot System, 12-well plates were placed in the frame of the Robot. ( D ) Addition of GFP Lentivirus to Cells, 70, 100, and 150° extractions were made from the well containing GFP Lentivirus (GFP LV) in a 12-well plate and added into wells 1, 2, and 3. ( E ) Incubation, after the robot performed the experiment, the 12-Well plate was placed in the incubator. ( F ) 70, 100, and 150° blocks were used for genetic transfer in human cells experiment programming. ( G ) 100°, and ( H ) 150° blocks that activate the pipette system are prepared in the experiment programming and perform the liquid withdrawal process. ( I ) Analysis of GFP protein expression in CRISPR.BOT modified transgenic Jurkat cell (Clone E6-1, TIB-152, ATCC) line using flow cytometry and ( J ) Fluorescence microscopy at 4X.

Techniques: Expressing, Cell Culture, Virus, Sample Prep, Incubation, Transferring, CRISPR, Modification, Transgenic Assay, Flow Cytometry, Fluorescence, Microscopy

CRISPR gene modification and flow cytometry analysis. ( A ) Seeding of Jurkat Cells (Clone E6-1, TIB-152, ATCC) into 12 well plate at 5 × 10 4 cells/well. ( B ) virus sample preparation. gRNA 1,2,3 Lentivirus was placed in 12 well plate. ( C ) Integration of plates into the autonomous robot system, 12-well plates were placed in the frame of the Robot. ( D ) Addition of gRNA lentivirus to cells. The wells containing gRNA Lentivirus 1,2,3 in the 12-well plate were drawn at 100° and added to wells 1, 2, and 3 separately. ( E ) Incubation. After the robot performed the experiment, the plate was placed in the incubator. ( F ) flow cytometry analysis of GFP protein expression in transgenic human cells encoding CRISPR.BOT modified CRISPR guide RNAs. gRNA guide RNA, GFP green fluorescent protein.

Journal: Scientific Reports

Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications

doi: 10.1038/s41598-025-01655-2

Figure Lengend Snippet: CRISPR gene modification and flow cytometry analysis. ( A ) Seeding of Jurkat Cells (Clone E6-1, TIB-152, ATCC) into 12 well plate at 5 × 10 4 cells/well. ( B ) virus sample preparation. gRNA 1,2,3 Lentivirus was placed in 12 well plate. ( C ) Integration of plates into the autonomous robot system, 12-well plates were placed in the frame of the Robot. ( D ) Addition of gRNA lentivirus to cells. The wells containing gRNA Lentivirus 1,2,3 in the 12-well plate were drawn at 100° and added to wells 1, 2, and 3 separately. ( E ) Incubation. After the robot performed the experiment, the plate was placed in the incubator. ( F ) flow cytometry analysis of GFP protein expression in transgenic human cells encoding CRISPR.BOT modified CRISPR guide RNAs. gRNA guide RNA, GFP green fluorescent protein.

Techniques: CRISPR, Modification, Flow Cytometry, Virus, Sample Prep, Incubation, Expressing, Transgenic Assay

Single-cell sub-cloning procedures of genetically modified cells, GFP expression and cell viability in subcloned cells. ( A ) Precise microliter liquid intake performance trial studies with pipette system for CRISPR.BOT V2. ( B ) Seeding of Cells. CRISPR + gRNA 1-2-3 Jurkat Cells (Clone E6-1, TIB-152, ATCC) were cultivated at 5 × 10 4 cells/well in a 6-well. ( C ) Cell dilution. 10 µl of 3 different viruses in the wells of the 6-well plate were taken separately. 1 ml/500 cells were taken from the cell + RPMI mixture with a total volume of 1 ml and added onto 9 ml RMPI medium and placed in 6-well. ( D ) Integration of plates into the autonomous robot system. 6-well and 96-well plates were placed on the Robot’s frame. ( E ) Sub-cloning of cells into 96-well plate. CRISPR + gRNA 1-2-3 Jurkat Cells (Clone E6-1, TIB-152, ATCC) in the 6-well plate were pulled at 100° and added to a whole 96-well plate. ( F ) Incubation of the 96-Well plate. ( G ) CRISPR.BOT-based subcloning of the transgenic cells. ( H ) Shows the positive and negative rates of GFP expression in cells sorted in the experiment performed with CRISPR.BOT. The positive rate of GFP expression (more than 90%) is defined as “GFPhi”, while the negative rate (less than 10%) is called “GFPlo”. At the end of the experiment, the majority of the samples were sorted, resulting in a statistically significant result. Furthermore, some samples showed moderate GFP expression (between 10% and 90%), which is referred to as “GFPint”. ( I ) Shows the percentage of viability of subcloned cells and indicates that the statistical significance of these results was confirmed by unpaired t-test, * p < 0.05. Not significant, NS. µl, microliter; GFP green fluorescent protein, gRNA guide RNA. This indicates that the results are reliable and that the experiment was successful.

Journal: Scientific Reports

Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications

doi: 10.1038/s41598-025-01655-2

Figure Lengend Snippet: Single-cell sub-cloning procedures of genetically modified cells, GFP expression and cell viability in subcloned cells. ( A ) Precise microliter liquid intake performance trial studies with pipette system for CRISPR.BOT V2. ( B ) Seeding of Cells. CRISPR + gRNA 1-2-3 Jurkat Cells (Clone E6-1, TIB-152, ATCC) were cultivated at 5 × 10 4 cells/well in a 6-well. ( C ) Cell dilution. 10 µl of 3 different viruses in the wells of the 6-well plate were taken separately. 1 ml/500 cells were taken from the cell + RPMI mixture with a total volume of 1 ml and added onto 9 ml RMPI medium and placed in 6-well. ( D ) Integration of plates into the autonomous robot system. 6-well and 96-well plates were placed on the Robot’s frame. ( E ) Sub-cloning of cells into 96-well plate. CRISPR + gRNA 1-2-3 Jurkat Cells (Clone E6-1, TIB-152, ATCC) in the 6-well plate were pulled at 100° and added to a whole 96-well plate. ( F ) Incubation of the 96-Well plate. ( G ) CRISPR.BOT-based subcloning of the transgenic cells. ( H ) Shows the positive and negative rates of GFP expression in cells sorted in the experiment performed with CRISPR.BOT. The positive rate of GFP expression (more than 90%) is defined as “GFPhi”, while the negative rate (less than 10%) is called “GFPlo”. At the end of the experiment, the majority of the samples were sorted, resulting in a statistically significant result. Furthermore, some samples showed moderate GFP expression (between 10% and 90%), which is referred to as “GFPint”. ( I ) Shows the percentage of viability of subcloned cells and indicates that the statistical significance of these results was confirmed by unpaired t-test, * p < 0.05. Not significant, NS. µl, microliter; GFP green fluorescent protein, gRNA guide RNA. This indicates that the results are reliable and that the experiment was successful.

Techniques: Subcloning, Genetically Modified, Expressing, Transferring, CRISPR, Incubation, Transgenic Assay

CRISPR.BOT autonomous systems. Experimental approaches that can be performed with CRISPR.BOT autonomous systems.

Journal: Scientific Reports

Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications

doi: 10.1038/s41598-025-01655-2

Figure Lengend Snippet: CRISPR.BOT autonomous systems. Experimental approaches that can be performed with CRISPR.BOT autonomous systems.

Techniques: CRISPR

CRISPR.BOT closed the laboratory automation system. Development of CRISPR.BOT closed laboratory autonomous systems and their integration with laboratory analyzer (RT-PCR, ELISA Plate Reader, Flow Cytometry, Incubator, and Thermal Cycler). RT-PCR real time-PCR.

Journal: Scientific Reports

Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications

doi: 10.1038/s41598-025-01655-2

Figure Lengend Snippet: CRISPR.BOT closed the laboratory automation system. Development of CRISPR.BOT closed laboratory autonomous systems and their integration with laboratory analyzer (RT-PCR, ELISA Plate Reader, Flow Cytometry, Incubator, and Thermal Cycler). RT-PCR real time-PCR.

Techniques: CRISPR, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Real-time Polymerase Chain Reaction

(A) Original and new designs of gRNA-markers for pMAGIC and nMAGIC. (B) Comparison of clone frequency in larval sensory neurons between two gRNA designs. The number represents clones between A1 and A7 segments on one side of each larva. n = larvae number: tgFE (n=10), Qtg2.1 (n=10). (C-E) Labeling of hemocytes in whole 3 rd instar larvae by pxn Gal4 >CD4-tdTom alone (C) or together with ubi-Gal80 (D) or tub-Gal80 (E). The panels on the right show enlarged views of the boxed regions. (F) Designs of Gal80 variants tested in pMAGIC gRNA-markers. (G) The brightness of epidermal clones labeled by pMAGIC gRNA-markers. n = image numbers: gRNA-40D2-uH (n = 32), gRNA-40D2-uDEH (n = 31), gRNA-42A4-uDEH (n = 52), gRNA-42A4-tDEH (n = 39), gRNA-42A4-tDES (n = 38). (H) The brightness of neuronal clones labeled by pMAGIC gRNA-markers. n = neuron numbers: gRNA-40D2-uH (n = 16), gRNA-40D2-uDEH (n = 16), gRNA-42A4-uDEH (n = 16), gRNA-42A4-tDEH (n = 15), gRNA-42A4-tDES (n = 16). (I) Portion of a larval wing disc containing nMAGIC clones visualized by nlsBFP. (J and J’) Portion of a wing disc containing nMAGIC clones labeled by cytosolic BFP (J) and HA staining (J’). (K) Epidermal clones on the larva body wall labeled by nlsBFP. (L) Epidermal clones visualized by cytosolic BFP. In all plots, black bar, mean; red bar, SD; AU, arbitrary unit. Student’s t-test in (B); one-way analysis of variance (ANOVA) and Tukey’s honest significant difference (HSD) test in (G) and (H). *p≤0.05, **p≤0.01, ***p≤0.001, ns, not significance. For (C-E), scale bar, 300 µm. For (I-M), scale bar, 100 µm.

Journal: bioRxiv

Article Title: A genome-wide MAGIC kit for recombinase-independent mosaic analysis in Drosophila

doi: 10.1101/2025.06.30.662354

Figure Lengend Snippet: (A) Original and new designs of gRNA-markers for pMAGIC and nMAGIC. (B) Comparison of clone frequency in larval sensory neurons between two gRNA designs. The number represents clones between A1 and A7 segments on one side of each larva. n = larvae number: tgFE (n=10), Qtg2.1 (n=10). (C-E) Labeling of hemocytes in whole 3 rd instar larvae by pxn Gal4 >CD4-tdTom alone (C) or together with ubi-Gal80 (D) or tub-Gal80 (E). The panels on the right show enlarged views of the boxed regions. (F) Designs of Gal80 variants tested in pMAGIC gRNA-markers. (G) The brightness of epidermal clones labeled by pMAGIC gRNA-markers. n = image numbers: gRNA-40D2-uH (n = 32), gRNA-40D2-uDEH (n = 31), gRNA-42A4-uDEH (n = 52), gRNA-42A4-tDEH (n = 39), gRNA-42A4-tDES (n = 38). (H) The brightness of neuronal clones labeled by pMAGIC gRNA-markers. n = neuron numbers: gRNA-40D2-uH (n = 16), gRNA-40D2-uDEH (n = 16), gRNA-42A4-uDEH (n = 16), gRNA-42A4-tDEH (n = 15), gRNA-42A4-tDES (n = 16). (I) Portion of a larval wing disc containing nMAGIC clones visualized by nlsBFP. (J and J’) Portion of a wing disc containing nMAGIC clones labeled by cytosolic BFP (J) and HA staining (J’). (K) Epidermal clones on the larva body wall labeled by nlsBFP. (L) Epidermal clones visualized by cytosolic BFP. In all plots, black bar, mean; red bar, SD; AU, arbitrary unit. Student’s t-test in (B); one-way analysis of variance (ANOVA) and Tukey’s honest significant difference (HSD) test in (G) and (H). *p≤0.05, **p≤0.01, ***p≤0.001, ns, not significance. For (C-E), scale bar, 300 µm. For (I-M), scale bar, 100 µm.

Article Snippet: The PCR product was then assembled with SapI-digested gRNA cloning vectors using NEBuilder DNA Assembly.

Techniques: Comparison, Clone Assay, Labeling, Staining

(A) Scheme of gRNA-marker insertion sites and target sites on Drosophila chromosomes. (B) Comparison of clone frequencies of all pMAGIC gRNA-markers in larval sensory neurons, clones are labeled using RabX4-Gal4 UAS-MApHs (for Chromosome X, II and IV) or 21-7-Gal4 UAS-MApHs (for Chromosome III). n = larvae number: X2 (n = 10), 20F2 (n = 10), 20F1(n = 10), 40D2 (n = 20), 40D4 (n = 10), 40E1 (n = 10), 41F9 (n = 20), 41F11 (n = 10), 42A4 (n = 10), 80C1 (n = 20), 80C2 (n = 14), 80F5 (n = 15), 81F (n = 10), 82A4 (n = 10), 82C3 (n = 10), 101F1a (n = 10), 101F1b (n = 10), 101F1c (n = 10). (C) Comparison of clone areas in larval wing discs labeled by nMAGIC gRNA-markers on 2R. n = wing disc number: 41F9 (n = 14), 41F11 (n = 16), 42A4 (n = 15). (D and E) Neuronal clones in the central part of the adult brain induced by pMAGIC gRNA-markers gRNA-40D2 (D) and gRNA-40E1 (E). In all plots, black bar, mean; red bar, SD. One-way ANOVA and Tukey’s HSD test. *p≤0.05, **p≤0.01, ***p≤0.001, ns, not significance. For (D) and (E), scale bar 100 µm.

Journal: bioRxiv

Article Title: A genome-wide MAGIC kit for recombinase-independent mosaic analysis in Drosophila

doi: 10.1101/2025.06.30.662354

Figure Lengend Snippet: (A) Scheme of gRNA-marker insertion sites and target sites on Drosophila chromosomes. (B) Comparison of clone frequencies of all pMAGIC gRNA-markers in larval sensory neurons, clones are labeled using RabX4-Gal4 UAS-MApHs (for Chromosome X, II and IV) or 21-7-Gal4 UAS-MApHs (for Chromosome III). n = larvae number: X2 (n = 10), 20F2 (n = 10), 20F1(n = 10), 40D2 (n = 20), 40D4 (n = 10), 40E1 (n = 10), 41F9 (n = 20), 41F11 (n = 10), 42A4 (n = 10), 80C1 (n = 20), 80C2 (n = 14), 80F5 (n = 15), 81F (n = 10), 82A4 (n = 10), 82C3 (n = 10), 101F1a (n = 10), 101F1b (n = 10), 101F1c (n = 10). (C) Comparison of clone areas in larval wing discs labeled by nMAGIC gRNA-markers on 2R. n = wing disc number: 41F9 (n = 14), 41F11 (n = 16), 42A4 (n = 15). (D and E) Neuronal clones in the central part of the adult brain induced by pMAGIC gRNA-markers gRNA-40D2 (D) and gRNA-40E1 (E). In all plots, black bar, mean; red bar, SD. One-way ANOVA and Tukey’s HSD test. *p≤0.05, **p≤0.01, ***p≤0.001, ns, not significance. For (D) and (E), scale bar 100 µm.

Article Snippet: The PCR product was then assembled with SapI-digested gRNA cloning vectors using NEBuilder DNA Assembly.

Techniques: Marker, Comparison, Clone Assay, Labeling

Journal: bioRxiv

Article Title: A genome-wide MAGIC kit for recombinase-independent mosaic analysis in Drosophila

doi: 10.1101/2025.06.30.662354

Figure Lengend Snippet:

Article Snippet: The PCR product was then assembled with SapI-digested gRNA cloning vectors using NEBuilder DNA Assembly.

Techniques:

(A-F) pMAGIC clones induced in different tissues by vas-Cas9 gRNA-40D2(Gal80) and labeled by tub-Gal4 UAS-CD8-GFP (green). DAPI staining (white) shows all nuclei. (G) A pMAGIC epidermal clone on the larval body wall induced by zk-Cas9 gRNA-40D2(Gal80) and labeled by R38F11-Gal4 UAS-tdTom (green). Epidermal junctions are labeled by α-Catenin-GFP (white). (H) pMAGIC glia clones in the larval brain induced by gcm-Cas9 gRNA-40D2(Gal80) and labeled by repo-Gal4 UAS-CD8-GFP (green). Glial nuclei are labeled by Repo staining (white). (I) pMAGIC hemocyte clones induced by Act-Cas9 gRNA-40D2(Gal80) and labeled by pxn-Gal4 UAS-tdTom . For figure A, D-F, H, scale bar 100 µm. For figure B-C, G, scale bar 50 µm. For Figure I, scale bar 25 µm.

Journal: bioRxiv

Article Title: A genome-wide MAGIC kit for recombinase-independent mosaic analysis in Drosophila

doi: 10.1101/2025.06.30.662354

Figure Lengend Snippet: (A-F) pMAGIC clones induced in different tissues by vas-Cas9 gRNA-40D2(Gal80) and labeled by tub-Gal4 UAS-CD8-GFP (green). DAPI staining (white) shows all nuclei. (G) A pMAGIC epidermal clone on the larval body wall induced by zk-Cas9 gRNA-40D2(Gal80) and labeled by R38F11-Gal4 UAS-tdTom (green). Epidermal junctions are labeled by α-Catenin-GFP (white). (H) pMAGIC glia clones in the larval brain induced by gcm-Cas9 gRNA-40D2(Gal80) and labeled by repo-Gal4 UAS-CD8-GFP (green). Glial nuclei are labeled by Repo staining (white). (I) pMAGIC hemocyte clones induced by Act-Cas9 gRNA-40D2(Gal80) and labeled by pxn-Gal4 UAS-tdTom . For figure A, D-F, H, scale bar 100 µm. For figure B-C, G, scale bar 50 µm. For Figure I, scale bar 25 µm.

Article Snippet: The PCR product was then assembled with SapI-digested gRNA cloning vectors using NEBuilder DNA Assembly.

Techniques: Clone Assay, Labeling, Staining

(A-A”) pMAGIC clones of VGlut 1 mutation in motor neurons at the neuromuscular junction. Clones were induced by zk-Cas9 gRNA-40D2(Gal80) and labeled by tub-Gal4 UAS-CD8-GFP ., The loss of VGlut is confirmed by VGlut staining. The mutant clones are outlined in (A”). (B-B”) A pMAGIC clone of brp d09839 mutation in a motor neuron at the neuromuscular junction. Clones were induced by zk-Cas9 gRNA-42A4(Gal80) and labeled by tub-Gal4 UAS-CD8-GFP . The loss of Brp is confirmed by Brp staining. The mutant clone is outlined in (B”). In both experiments, HRP staining shows all axons. Scale bars, 10 µm.

Journal: bioRxiv

Article Title: A genome-wide MAGIC kit for recombinase-independent mosaic analysis in Drosophila

doi: 10.1101/2025.06.30.662354

Figure Lengend Snippet: (A-A”) pMAGIC clones of VGlut 1 mutation in motor neurons at the neuromuscular junction. Clones were induced by zk-Cas9 gRNA-40D2(Gal80) and labeled by tub-Gal4 UAS-CD8-GFP ., The loss of VGlut is confirmed by VGlut staining. The mutant clones are outlined in (A”). (B-B”) A pMAGIC clone of brp d09839 mutation in a motor neuron at the neuromuscular junction. Clones were induced by zk-Cas9 gRNA-42A4(Gal80) and labeled by tub-Gal4 UAS-CD8-GFP . The loss of Brp is confirmed by Brp staining. The mutant clone is outlined in (B”). In both experiments, HRP staining shows all axons. Scale bars, 10 µm.

Article Snippet: The PCR product was then assembled with SapI-digested gRNA cloning vectors using NEBuilder DNA Assembly.

Techniques: Clone Assay, Mutagenesis, Labeling, Staining

(A) A WT pMAGIC class IV da neuron clone exhibiting complete dendrite pruning at 16 hours APF. (B-D) pMAGIC clones of EcR M554fs mutation in da neurons imaged at 16 hours APF, exhibiting the lack of pruning (B and D) or apoptosis (C). In (A-D), the clones were induced by zk-cas9 with gRNA-41F9(Gal80) and labeled by RabX4-Gal4 UAS-MApHS . Neuronal cell bodies are indicated by arrows. MApHS contains pHluorin and tdTom , but only tdTom signals are shown. The signals in epidermal cells (A) were due to engulfment of pruned dendrites by epidermal cells . (E and F) WT (E) and Df(4)ED6380 (F) pMAGIC clones in C4da neurons induced by zk-cas9 gRNA-101Fc(Gal80) and labeled by RabX4-Gal4 UAS-MApHS . Only tdTom signals are shown. (G) Normalized dendrite length of WT clones and deficiency clones. Black bar, mean; red bar, SD. Student’s t-test. ***p≤0.001. (H) Scheme for interspecific crosses between D. melanogaster ( D.m ) and D. simulans ( D.s ). (I and J) Wing discs from male (I) and female (J) progeny carrying clones. Scale bars, 50 µm.

Journal: bioRxiv

Article Title: A genome-wide MAGIC kit for recombinase-independent mosaic analysis in Drosophila

doi: 10.1101/2025.06.30.662354

Figure Lengend Snippet: (A) A WT pMAGIC class IV da neuron clone exhibiting complete dendrite pruning at 16 hours APF. (B-D) pMAGIC clones of EcR M554fs mutation in da neurons imaged at 16 hours APF, exhibiting the lack of pruning (B and D) or apoptosis (C). In (A-D), the clones were induced by zk-cas9 with gRNA-41F9(Gal80) and labeled by RabX4-Gal4 UAS-MApHS . Neuronal cell bodies are indicated by arrows. MApHS contains pHluorin and tdTom , but only tdTom signals are shown. The signals in epidermal cells (A) were due to engulfment of pruned dendrites by epidermal cells . (E and F) WT (E) and Df(4)ED6380 (F) pMAGIC clones in C4da neurons induced by zk-cas9 gRNA-101Fc(Gal80) and labeled by RabX4-Gal4 UAS-MApHS . Only tdTom signals are shown. (G) Normalized dendrite length of WT clones and deficiency clones. Black bar, mean; red bar, SD. Student’s t-test. ***p≤0.001. (H) Scheme for interspecific crosses between D. melanogaster ( D.m ) and D. simulans ( D.s ). (I and J) Wing discs from male (I) and female (J) progeny carrying clones. Scale bars, 50 µm.

Article Snippet: The PCR product was then assembled with SapI-digested gRNA cloning vectors using NEBuilder DNA Assembly.

Techniques: Clone Assay, Mutagenesis, Labeling

(A-B) Representative epidermal (A) and wing disc (B) images showing uneven expression of gRNA-101F1c(BFP) inserted at attP 102D on the 4 th chromosome. Yellow arrowheads point to cells lacking BFP expression. Scale bars, 50 µm. (C) Frequency of labeled neurons by indicated gRNA(Gal80) in the absence and presence of Cas9. The zk-Cas9 dataset is the same as that for chromosome 4 in . Black bar, mean; red bar, SD. One-way ANOVA and HSD test. ***p≤0.001.

Journal: bioRxiv

Article Title: A genome-wide MAGIC kit for recombinase-independent mosaic analysis in Drosophila

doi: 10.1101/2025.06.30.662354

Figure Lengend Snippet: (A-B) Representative epidermal (A) and wing disc (B) images showing uneven expression of gRNA-101F1c(BFP) inserted at attP 102D on the 4 th chromosome. Yellow arrowheads point to cells lacking BFP expression. Scale bars, 50 µm. (C) Frequency of labeled neurons by indicated gRNA(Gal80) in the absence and presence of Cas9. The zk-Cas9 dataset is the same as that for chromosome 4 in . Black bar, mean; red bar, SD. One-way ANOVA and HSD test. ***p≤0.001.

Article Snippet: The PCR product was then assembled with SapI-digested gRNA cloning vectors using NEBuilder DNA Assembly.

Techniques: Expressing, Labeling

Review

Structured Review

Images

Similar Products

Image Search Results