RS-1 is a RAD51 activator, and also increases CRISPR/Cas9-mediated knock-in efficiencies.

RAD51^[1], CRISPR/Cas9^[2]

RS-1 is a RAD51 activator, stimulating binding of hRAD51 to DNA with K_d ranging from 48 nM to 107 nM in the presence of ATP or ADP and in the absence of a nucleotide cofactor, and such an effect is not via inhibiting its ATPase activity. RS-1 (20 μM) affects the length and helical pitch of hRAD51 protein-DNA complexes. RS-1 (0, 1, 5, 10, 15, 20, and 25 μM) stimulates strand assimilation activity of hRAD51. RS-1 (7.5 μM) promotes resistance of human cells to cross-linking chemotherapy^[1]. RS-1 (0, 7.5, 15 μM) increases Cas9-mediated knock-in efficiencies in rabbit embryos^[2].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

524.23

C₂₀H₁₆Br₂N₂O₃S

312756-74-4

Room temperature in continental US; may vary elsewhere.

Powder	-20°C	3 years
	4°C	2 years
In solvent	-80°C	6 months
	-20°C	1 month

In Vitro: 

DMSO : ≥ 100 mg/mL (190.76 mM)

* “≥” means soluble, but saturation unknown.

*

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

In Vivo:

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 3 mg/mL (5.72 mM); Clear solution

  此方案可获得 ≥ 3 mg/mL (5.72 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 30.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 3 mg/mL (5.72 mM); Clear solution

  此方案可获得 ≥ 3 mg/mL (5.72 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 30.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Jayathilaka K, et al. A chemical compound that stimulates the human homologous recombination protein RAD51. Proc Natl Acad Sci U S A. 2008 Oct 14;105(41):15848-53.

  [2]. Song J, et al. RS-1 enhances CRISPR/Cas9- and TALEN-mediated knock-in efficiency. Nat Commun. 2016 Jan 28;7:10548.

Briefly, 15 μL reaction volumes include a DNA strand exchange protein (0.8 μM) that is preincubated for 5 min at 37°C with 1 μM (nucleotide concentration) ³²P-labeled oligonucleotide 306.7 in a reaction buffer containing 20 mM Hepes (pH 7.5), 1 mM DTT, 2 mM nucleotide cofactor, and 1 mM MgCl₂ and various concentrations of RS-1. For experimental buffer conditions that included calcium, 1 mM CaCl₂ is present in addition to (in the case of hRAD51) or in the place of (in the case of RecA and scRAD51) the 1 mM MgCl₂. Conditions with scRAD51 additionally contains 110 nM scRAD54. After this initial binding reaction, 10 μL of 19.75 μM (base pair concentration) supercoiled homologuecontaining target plasmid DNA (pRS306) is next added along with sufficient magnesium acetate to give a final concentration of 10 mM^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Jayathilaka K, et al. A chemical compound that stimulates the human homologous recombination protein RAD51. Proc Natl Acad Sci U S A. 2008 Oct 14;105(41):15848-53.

  [2]. Song J, et al. RS-1 enhances CRISPR/Cas9- and TALEN-mediated knock-in efficiency. Nat Commun. 2016 Jan 28;7:10548.